[Tumor necrosis factor in the ejaculate as an indicator of reduced fertility].

INTRODUCTION: Pro-inflammatory cytokine - tumor necrosis factor-alpha (TNF) is one of the components of the seminal plasma proteome; its meaning has not been definitively revealed. A comparative analysis of the concentration of this protein in the blood serum and in the ejaculate and changes in its level in the semen of men with infertility is f scientific interest. THE PURPOSE OF THE STUDY: determination of TNF- level in the blood serum and seminal plasma of healthy men and patients with reduced fertility. MATERIALS AND METHODS: 70 men of reproductive age with azoospermia (main group, n=18), with oligoastenozoospermia (comparison group, n=18) and with normal spermogram parameters (control group, n=34) were examined. The ejaculate was examined using an SQA-V semen analyzer (MES, Israel). In seminal plasma samples, the concentration of TNF was determined using the alpha-TNF-ELISA-BEST test system (A-8756, Vector-Best LL, Russia). RESULTS: The concentration of TNF- in blood serum had a significant variation (CV=85.31%) and amounted to 2.75+/-2.18 pg/ml, which is 2.55 times lower than the same indicator in seminal plasma (7.01+/-5.98 pg/ml, CV=126.15%, p<0.00001). When comparing the content of TNF- in seminal plasma, significant differences were found in the examined patients (Kruskal-Wallis test H=24.75991; p<0.00001). Pairwise comparison revealed a statistically significant difference in the level of TNF- in seminal plasma between the comparison and control groups (p2-3=0.000023), as well as between the main group and the comparison group (p1-2=0.000043); there were no significant differences between the main and control groups (p>0.05). When determining the content of TNF- in the blood serum, there was no statistically significant difference between the groups (p>0.05). There were no correlations between the concentration of TNF- in blood serum and in seminal plasma (R=0.295374), and the total number of spermatozoa in the ejaculate (R=-0.027945); and the concentration of spermatozoa in the ejaculate (R=-0.042902). DISCUSSION: It is unlikely that TNF crosses into seminal plasma from serum against a concentration gradient. It is most likely that TNF is produced locally in the organs of the reproductive system by resident immune cells or cells involved in spermatogenesis. An increased content of TNF- in seminal plasma in patients of the comparison group may indicate the presence of an inflammatory process in the reproductive system and a reduced fertility of the ejaculate. CONCLUSION: The physiological role of TNF in sperm, its sources in the organs of the male reproductive system, and the pathogenetic mechanisms of the participation of the TNF in pathological processes in male reproductive system still remain unclear. All this justifies the need for further study of the TNF level in seminal plasma in normal conditions and in diseases of the urogenital tract in men.

Comparative Evaluation of the Parameters of Sperm Apoptosis of Young and Middle-Aged Men by Flow Cytometry.

A comparative analysis of the parameters characterizing sperm apoptosis of young (27-42 years) and middle-aged (44-51 years) men was performed by flow cytometry. Irrespective of age, activity of caspase-3 and p53-mediated controlling the transmission of apoptogenic signal transmission in gametes remained stable with the formation of germ cells with delayed (p<0.05) cell death according to the Annexin V-FITC+PI- criterion (predominantly in middle-aged men). Inhibition of the transmission of a proapoptogenic stimulus mediated by membrane cell death receptors (FAS) was also observed in this group. Comparison of indicators of sperm apoptosis showed age-related features of cell death, in particular, inhibition of membrane reception triggering FAS-dependent apoptosis, which is associated with insufficient phosphatidylserine production in middle-aged men, excessive life cycle duration, and aging of spermatozoa.

[Fatty acid-binding protein in the serum and sperm].

INTRODUCTION: a composition of seminal plasma reflects a state of reproductive organs that are involved in sperm production. AIM: to study a concentration of fatty acid-binding protein (FABP) in blood serum and sperm samples with normal and altered characteristics. MATERIALS AND METHODS: a total of 82 men with mean age of 33.1+/-4.7 years old who were examined to clarify the cause of infertility were included in the study. The main group (n=21) consisted of men with oligozoospermia, while control group (n=33) consisted of healthy men with normal sperm analysis. In addition, patients with high viscosity of sperm were included in the comparison group (n=28). There were no changes in complete blood count and biochemical blood panel in all participants. The content of FABP in blood serum and seminal plasma was determined. RESULTS: the average content of FABP in seminal plasma was 1.347+/-0.26 ng/ml and exceeded the serum concentration of this compound (p<0.000001), which was 0.305+/-0.193 ng/ml. The concentration in seminal plasma did not depend on its serum concentration (R=0,068194), as well as the sperm volume and viscosity, but it was correlated with sperm concentration (R=0,66387). The concentration of FABP in seminal plasma was highest in the control group (1.47+/-0.33 ng / ml) and it was higher compared to the main group (1.22+/-0.09 ng/ml; p=0.000057) and the comparison group (1.29+/-0.19 ng/ml; p=0.010822). There was no difference between groups in serum concentration of FABP (p=0.9814). CONCLUSION: the obtained data suggest that a reduced level of FABP in seminal plasma can be considered as unfavorable criterion indicating a reduced fertility.

Analysis of the Role of Carriership of Polymorphic Genotypes of ESR1, eNOS, and APOE4 Genes in the Development of Arterial Hypertension in Men.

We studied the role of the carrier status for polymorphic loci of genes encoding estrogen receptors (ESR1), endothelial NO synthase (eNOS), and apolipoprotein E (APOE4) and products of their expression nitrogen oxide (NO) and apolipoprotein (ApoE) in the development of arterial hypertension in men. Conventionally healthy volunteers and 149 men with clinical manifestations of stage I-II arterial hypertension were examined. In men with arterial hypertension, the frequency of minor allele A of ESR1 gene was higher (27.5 vs. 9.5% in the reference group; χ2=4.43, p=0.04). The level of NO in the peripheral blood was also higher in the main group (χ2=3.93, p=0.047). The increase in NO concentration did not depend on the presence of polymorphic genotypes (GG and GT) of eNOS gene, but the decrease in ApoE level in blood serum was associated with TC genotype of APOE4 gene (p=0.04). Our results suggest that minor allele A of ESR1 gene is associated with the development of arterial hypertension in men. Reduced content of ApoE in blood serum of men with arterial hypertension was associated with APOE4 gene polymorphism. However, increased level of NO did not depend on polymorphic genotypes GG and GT of eNOS gene. These polymorphisms are of specific interest as additional markers of genetic predisposition to the development of arterial hypertension in middle-age men.

[The NT-proBNP in saliva and blood serum in norm and under periodontitis.]

The concentration of NT-proBNP in saliva and blood serum was studied. The study included 58 individuals divided in two groups. The main group (group I) included 34 patients with chronic generalized periodontitis. The control group (group II), comparable in age and gender, included 24 examined individuals without signs of inflammatory diseases of oral cavity. The concentration of NT-proBNP in saliva and blood serum was detected by technique of solid-phase enzyme immunoassay using the test-system "NT-proBNP-ELISABEST" (Vector-Best, Russia). The concentration of NT-proBNP in saliva of patients of main group reliably exceeded its concentration in saliva of the examined from control group. The median and interquartile range made up to correspondingly 178.5 (128-253) pg / ml in main group and 105 (72.5 - 144.5) pg, respectively / ml in control group (p = 0.02). The comparison of concentration of NT-proBNP in blood serum samples established no reliable differences (p = 0,27). The evaluation of the relationship between content of NT-proBNP in saliva and blood serum demonstrated absence of close linear correlation dependence between the concentration of this compound in blood serum and saliva both in the main group (R = 0.143, p = 0.419) and the control group (R = 0.178; p = 0.405), and for all the examined as well (R = 0.252, p = 0.056). The higher concentration of NT-proBNP in saliva in comparison with blood serum can be explained by formation of proteolysis products cross-reacting in the used system.

[Ejaculate and serum procalcitonin levels in healthy men and men with oligoasthenozoospermia].

INTRODUCTION: Seminal plasma composition reflects the activity of reproductive organs involved in the semen production. AIM: To study procalcitonin concentrations in serum and semen samples of healthy men and men with oligoasthenozoospermia. METHOD: s .The study included 88 men, who were scheduled for diagnostic evaluation to establish the cause of infertile marriages. The study group comprised 40 men with oligoasthenozoospermia, the comparison group included 48 men with normal sperm concentration. Laboratory testing of all participants revealed no abnormal findings in blood count, blood chemistry studies and urinalysis. RESULTS: Mean seminal plasma procalcitonin level in the study subjects (n=87) was 0,349+/-0,370 ng/ml being about 10 times higher than its serum level, which was 0.037+/-0.027 ng/ml (p<0.000001). In the study group, seminal plasma PCT concentration was significantly greater than in the control group (p=0.0095), while the serum procalcitonin levels in all participants were almost identical (p=0.605). There were no statistically significant correlations between the procalcitonin levels and spermatozoa concentration, total count and ejaculate volume. CONCLUSIONS: The findings suggest that elevated levels of procalcitonin in seminal plasma can be regarded as an unfavorable prognostic factor, indicating the reduced ejaculate fertility. Further studies seem warranted, specifically considering the role and source of procalcitonin production in sperm.

Patient-derived xenotransplants can recapitulate the genetic driver landscape of acute leukemias.

Genomic studies have identified recurrent somatic mutations in acute leukemias. However, current murine models do not sufficiently encompass the genomic complexity of human leukemias. To develop preclinical models, we transplanted 160 samples from patients with acute leukemia (acute myeloid leukemia, mixed lineage leukemia, B-cell acute lymphoblastic leukemia, T-cell ALL) into immunodeficient mice. Of these, 119 engrafted with expected immunophenotype. Targeted sequencing of 374 genes and 265 frequently rearranged RNAs detected recurrent and novel genetic lesions in 48 paired primary tumor (PT) and patient-derived xenotransplant (PDX) samples. Overall, the frequencies of 274 somatic variant alleles correlated between PT and PDX samples, although the data were highly variable for variant alleles present at 0-10%. Seventeen percent of variant alleles were detected in either PT or PDX samples only. Based on variant allele frequency changes, 24 PT-PDX pairs were classified as concordant while the other 24 pairs showed various degree of clonal discordance. There was no correlation of clonal concordance with clinical parameters of diseases. Significantly more bone marrow samples than peripheral blood samples engrafted discordantly. These data demonstrate the utility of developing PDX banks for modeling human leukemia, and emphasize the importance of genomic profiling of PDX and patient samples to ensure concordance before performing mechanistic or therapeutic studies.

[The significance of interleukin-6 and polymorphism of its gene (C174G) under viral, alcoholic and mixed cirrhosis of liver].

The study was carried out to evaluate significance of interleukin-6 (IL-6) and polymorphism of gene IL-6 (C174G) under liver cirrhosis of viral, alcoholic and mixed etiology. The blood of patients with liver cirrhosis of viral (n=48), alcoholic (n=11) and mixed (n=10) genesis was used for analyzing indices, level of IL-6 and polymorphism of gene IL-6 (C174G). In patients with liver cirrhosis independently of etiology increasing of concentration of IL-6 was established as compared with control group (p<0.001). The gene reliably correlated with marker of cytolysis (r=0.32; p=0.009), direct bilirubin (r=0.26; p=0.04), gamma-glutamyltranspeptidase (r=0.36; p=0.004), albumin (r=0.59; p<0.001) and degree of severity of liver cirrhosis according the Child-Pugh scale (r=0.76; r<0.001). The concentration of IL-6 was significantly higher in patients with alcoholic liver cirrhosis tahn in patients with viral and mixed genesis of disease (p=0.02 and p=0.02 correspondingly). The analysis of prevalence of genotypes and alleles of polymorphism of gene IL-6 (C174G) in groups of donors and patients with liver cirrhosis in the Permskii Kraii established no reliable differences in rate of occurrence of minor allele G and its association with production of IL-6 in examined cohorts (p=0.82 and p=0.29 correspondingly). This result indicates absence of relationship of probability of immune disorders and their genetic determination. The development of liver cirrhosis independently of etiologic factor is characterized by hyper-production of anti-inflammatory cytokine reflecting severity of lesion of hepatocytes, intensity of inflammatory process and progression of disease. The more intensive production of IL-6 was observed in case of alcoholic genesis of disease that conditioned fast progression of cirrhosis.

[THE INFLUENCE OF TUMOR NECROSIS FACTOR ALPHA AND POLYMORPHISM OF ITS GENE (RS1800629) ON THE SEVERITY AND PROGRESSION OF CHRONIC HEPATITIS AND ULCERATIVE COLITIS].

AIM OF THE STUDY: To investigate the effect of generation of tumor necrosis factor-alpha (TNFα) and the importance of TNFα(rs1 800629) gene polymorphism in the progression of chronic hepatitis C (CHC) and ulcerative colitis (UC). MATERIAL AND METHODS: The study involved 90 patients with chronic hepatitis C, 50 patients with UC and 50 healthy donors. The blood concentrations TNFα and TNFα gene polymorphism (rs1800629) were evaluated. RESULTS: TNFα levels in the blood in patients with chronic hepatitis C were increased compared with the control group and correlated with the severity of Cytolysis and fibrosis (r = 0.34, p = 0.02). At slow speed the formation of liver fibrosis TNFα amounted to 1.5 (0.9-2.8) pg/mI, with a fast speed--2.3(1.4-8.2) pg/mI (p = 0.006). Patients with UC at 3-4 degrees endo- scopic activity production of TNFα reached 6.5 (7-9) pg/mI, which was significantly higher than the value obtained at 1-2 degrees endoscopic activity--0.25(0-0.8) pg/ml (p = 0.001). The allelic variations of TNFα in groups of patients with CHC at different rates forming LF statistically differences were not found. The allele, associated with severe progressive course of UC and increased production of TNFα--A risk allele and genotype GA TNFα, associated with a slow progression of UC- "protective" G allele and genotype GG TNFα gene were determined. CONCLUSION: Determining the level of TNFα allows to evaluate the severity of liver disease, heaviness and progression of liver fibrosis speed in CHC, and the severity of inflammation in the intestinal mucosa in UC. The presence of the allele A of TNFo(rs1800629) is a predictor of severe and progression of UC. Determining genetic polymorphism TNFα in patients with UC may be an additional factor to assess the prognosis of the disease.

[FEATURES OF THE IMMUNE GENETIC PARAMETERS IN WORKERS IN NON-FERROUS METAL INDUSTRY].

There was performed a comparative analysis of immunogenetic indices in non-ferrous metallurgy employees under the exposure to different combinations of harmful occupational factors. The combined effect of chlorine and vanadium fumes, noise, overall vibration appeared to be associated with the gene polymorphism of cytokine regulation--VEGF and TNF (p < 0.05). In workers the combination offactors such as dust containing silicon dioxide, noise, elevated environmental thermal load was associated with cytochrome p450 gene polymorphism, allele variation ofwhich is formed owing to the homozygous genotype. At the same time there was observed an excess production of specific antibodies to vanadium and silicon, significantly differed from that of the indices in the reference group. There are proposed genetic (CYP1A1, VEGF TNFalfa) and immunological (IgG to vanadium and silicon) indices as markers of susceptibility and effect in health risk assessment of different combinations of harmful occupational factors, which will allow to increase the availability of laboratory control during surveillance activities at the objects.

[Metabolic disorder and polymorphism of the genes encoding for beta-2-adrenergic receptor and apolipoproteins B in chronic hepatitis C and non-alcoholic fatty liver diseases].

UNLABELLED: The aim of the study was to evaluate the relationship between metabolic disorders and polymorphic variants of the genes encoding for beta-2-adrenergic receptor (ADRB2 (rs1042713) and apolipoproteins B (ApoB(rs5742904) in patients with chronic hepatitis C (CHC) depending on virus genotype and in patients with non-alcoholic fatty liver diseases (NAFLD) and concomitant metabolic syndrome (MS). MATERIALS AND METHODS: The study included 96 patients with CHC (51 with genotype 1 or 2 of hepatitis virus and 45 with genotype 3), 70 patients with NAFLD and MS, 51 healthy donors (controls). Gene polymorphism was studied by PCR (Sintol, Moscow) with a Real-time CFX-96 amplifier (Bio-Rad Lab. Inc., USA). RESULTS: CHC patients regardless of genotype had hypertriglyceridemia with increased atherogenicity index and C-peptide level. Hyperleptinemia was most frequently associated with genotype 3, hyperinsulinemia and insulin resistance with genotypes 1 and 2. The viremia level positively correlated with leptin level (p-0.021) and HOMA-IR index (p=0.022) which suggests virus-induced inactivation of mechanisms of steatogenesis and insulin resistance. Leptin production in CHC was associate with activation of liver fibrosis as appears from elasticity index measured by fibroelastography (p-0.22). Almost all patients with NAFLD had disturbances of lipid and carbohydrate metabolism. Hepatic lesions in 30% of the patients with MS were accompanied by laboratory signs of steatohepatitis. Patients with CHC showed an increased frequency of minor A allele of the ADRB2 (rs1042713) gene (up to 40%; p=0.04) and pathological A/A homozygote (22%; p=0.04). The occurrence of A allele was associated with hyperleptinemia (p=0.019). In NAFLD patients, the occurrence of A allele was higher than in controls (41%; p=0.02) with 55% of the case being A/G heterozygotes (p=0.005). The occurrence of A allele was related to hyperinsilinism (p=0.036), BMI (p=0.0330, and C-peptide production (p=0.038). No difference between the frequency of genotypes and ApoB(rs5742904) gene alleles in the patients of both groups and controls was documented. It is concluded that ADRB2 (rs1042713) gene polymorphism is associated with an increased risk of metabolic disorders in CHC and especially NAFLD with MS that aggravates liver disturbances. Dyslipidemia and insulin resistance in CHC patients is stimulated by viral infection.

[The noninvasive evaluation of degree of expression of fibrosis of liver and significance of polymorphism of gene of hyaluronic acid under chronic hepatitis C].

The study was carries out to evaluate degree of expression of fibrosis, reparation processes in liver and value of polymorphism of gene of hyaluronic acid HASI (rs11084111) in progression of affection of liver in patients with chronic hepatitis C. The sampling included 100 patients with chronic hepatitis C. The control group included 83 healthy donors. The blood serum was tested to detect concentration of hyaluronic acid and alpha-fetoprotein. The stage of liver fibrosis (F) was evaluated by using ultrasound fibroflexography The polymorphism of gene (rs11084111) was analysed by polymerase chain reaction technique. In the group of patients with F1 the average concentration of hyaluronic acid in blood serum in 1.8 times surpassed this indicator in group with F0. The concentration of hyaluronic acid was almost 2 times higher under F3 as compared with F1-F2. This indicator permitted differentiating F3 and F4 which followed by activation of cytolysis and cholestasis in F1 and F3 and by increasing of level of alpha-fetoprotein at stages F1 and F4. The study detected no statistically significant difference between rates of genotypes and alleles of gene HASI (rs11084111) in groups of healthy patients and patients with chronic hepatitis C. The direct relationships are established between hyaluronic acid and markers of cytolysis, cholestasis, alpha-fetoprotein (p = 0.001), viral load (p = 0.003) liver elasticity index according fibroflexography data (p < 0.001) and fibrosis index (p < 0.001). The established relationships indicate association of hepatofibrosis with cytolysis, cholestasis, hepatocytes regeneration and virus activity. The hyaluronic acid permits to stratify minimal expressed fibrosis and also the transition of disease to the stage of cirrhosis.

[Catalase gene rs1001179 polymorphism and oxidative stress in patients with chronic hepatitis C and ulcerative colitis].

AIM: To study the rs1001179 polymorphism of the catalase (CAT) gene and to estimate the serum levels of the enzymes catalase and glutathione peroxidase (GP) in patients with chronic hepatitis C (CHC) and in those with ulcerative colitis (UC) in the Perm Territory. SUBJECTS AND METHODS: Ninety patients with reactivation-phase CHC and 50 patients with exacerbation-phase UC were examined. The serum levels of catalase and GP were determined and the polymorphic variants of the marker of CAT gene rs1001179 in the DNA isolated from whole blood were found in all the patients. RESULTS: In the CHC and UC groups, the levels of catalase and GP were found to be lower than that in apparently healthy individuals. Furthermore, both groups showed a direct correlation between the activities of the enzymes. In the patients with CHC and in those with UC, the spread of genotypes and alleles generally failed to virtually differ from that in the control group. The G/G genotype was prevalent in all the groups. In the patients with CHC, the minor A allele demonstrated a significant inverse correlation with the enzyme catalase (r = -0.16; p = 0.02) and GP (r = -0.13; p = 0.047). CONCLUSION: The lower serum levels of catalase and GP are indicative of oxidative stress in the patients with CHC or UC. In the patients with CHC, the significant correlation of the pathological rs1701179 A allele marker with the processes of synthesis of antioxidant enzymes may suggest that CAT gene polymorphism in the A/A homozygotes might affect the regulation mechanism involved in the antioxidant system in the liver.

[Immune and genetic markers revealed in women working in technical rubber goods production].

Medical examination covered female workers engaged into chemical production of technical rubber goods, under combined exposure to chemicals (nitrogen oxides, carbon oxide) and noise exceeding MAC by 3.7-27.5%. Findings are increased phagocytary activity, lower level of serum IgM and IgG, suppressed expression of CD25 and CD95 T-cell receptors, and slower apoptosis induced via Fas receptor--all those indicators associated with work conditions. The disorders revealed are actualized on a basis of genetic variability features of the females, that is characterized by prevalence of mutant allels of FAS and TNFR genes with minor homozygous phenotype.

[Justifying genetic and immune markers of efficiency and sensitivity under combined exposure to risk factors in mining industry workers].

The authors evaluated and justified immunologic and genetic markers under combined exposure to risk factors in mining industry workers. Analysis covered polymorphism features of 29 genes with variant alleles possibly participating in occupationally conditioned diseases formation and serving as sensitivity markers of these diseases risk. The genes association selected demonstrates reliably changed polymorphism vs. the reference group (SOD2 superoxidedismutase gene, ANKK1 dophamine receptor gene, SULT1A1 sulphtransaminase gene, MTHFR methylene tetrahydrofolate reductase gene, VEGF endothelial growth factor gene, TNF-alpha tumor necrosis factor gene). Under combined exposure to occupational hazards (sylvinite dust, noise) in mining industry, this association can serve as adequate marking complex of sensitivity to development of occupationally conditioned diseases. Increased-production of immune cytokine regulation markers: tumor necrosis factor and vascular endothelial growth factor. Genes SOD2, ANKK1, SULT1A1, VEGF, TNFalpha are recommended as sensitivity markers, and the coded cytokines (tumor necrosis factor and endothelial growth factor) are proposed as effect markers in evaluation of health risk for workers in mining industry.

[The relationship between content of malonic aldehyde, activity of glutathione peroxidase and polymorphism of gene GPX4 (718C/T) under chronic hepatitis C].

The study was carried out to evaluate concentration of malonic dialdehyde and activity of glutathione peroxidase in blood serum in their relationship with biochemical tests of functional conditions of liver; data of fibroelastography and polymorphism of gene GPX4 (718C/T) in patients with chronic hepatitis C. The sampling consisted of 100 patients with chronic hepatitis C. The control group included 80 healthy donors. The polymorphism of gene of glutathione peroxidase was analyzed with CFX-96 (Bio-Rad Laboratories, Inc., USA) amplifier using technique of polymerase chain reaction ("Sintol", Moscow). In patients with chronic hepatitis C a reliable increasing of concentration of malonic dialdehyde up to 3.2 times and decreasing of glutathione peroxidase up to 2.8 times was detected as compared with control group. The significant differences in rates of genotypes and alleles of gene GPX4 (718C/T) in patients with chronic hepatitis C and healthy persons were not detected. The malonic dialdehyde demonstrated direct reliable relationships with functional hepatic tests by activity of alanine aminotransferase, aspartate aminotransferase and γ-glutamyl transferase. The activity of glutathione peroxidase had reverse reliable correlations with alanine aminotransferase, aspartate aminotransferase and γ-glutamyl transferase (p=0.001, p=0.007, p=0.032) and with indicator of elasticity of liver according data of fibroelastography (r=-0.285, p=0.041) The minor allele T of gene GPX4 ()718C/T) reliably correlated with alanine aminotransferase, aspartate aminotransferase and malonic dialdehyde. The reverse relationship between allele T of gene GPX4 ()718C/T) and activity of glutathione peroxidase (r=-0.196, p=0.041) was established This occurrence indicates at negative effect of mutation in gene of glutathione peroxidase on functional activity of this enzyme. Under chronic hepatitis C, the activation of peroxidation of lipids and depression of activity of glutathione peroxidase are interrelated with severity of cytolysis, cholestasis, expression of hepatic fibrosis and polymorphism of gene GPX4 ()718C/T). Therefore, polymorphism of gene of glutathione peroxidase predisposes to more severe affection of liver and progression of chronic hepatitis C.

[Immunogenetic features of apoptosis in workers engaged in methanol production].

Disordered cellular death process with inhibition is seen during stress states including occupational activities associated with increased concentrations of methanol in biologic media. Workers exposed to methanol demonstrate prevalence of homozygous type of tumor necrosis factor (TNF) gene and heterozygous variant of cytochrome 450 gene, reliably increased vs reference group. Genetic changes in the main group are associated with reliable decrease in occurrence of transcription protein p53 and TNF receptor, and with altered ratio of intracellular apoptosis factors (bax and bcl-2) towards slower programmed cellular death in chemical production.

Cell of origin determines clinically relevant subtypes of MLL-rearranged AML.

Mixed lineage leukemia (MLL)-fusion proteins can induce acute myeloid leukemias (AMLs) from either hematopoietic stem cells (HSCs) or granulocyte-macrophage progenitors (GMPs), but it remains unclear whether the cell of origin influences the biology of the resultant leukemia. MLL-AF9-transduced single HSCs or GMPs could be continuously replated, but HSC-derived clones were more likely than GMP-derived clones to initiate AML in mice. Leukemia stem cells derived from either HSCs or GMPs had a similar immunophenotype consistent with a maturing myeloid cell (LGMP). Gene expression analyses demonstrated that LGMP inherited gene expression programs from the cell of origin including high-level Evi-1 expression in HSC-derived LGMP. The gene expression signature of LGMP derived from HSCs was enriched in poor prognosis human MLL-rearranged AML in three independent data sets. Moreover, global 5'-mC levels were elevated in HSC-derived leukemias as compared with GMP-derived leukemias. This mirrored a difference seen in 5'-mC between MLL-rearranged human leukemias that are either EVI1 positive or EVI1 negative. Finally, HSC-derived leukemias were more resistant to chemotherapy than GMP-derived leukemias. These data demonstrate that the cell of origin influences the gene expression profile, the epigenetic state and the drug response in AML, and that these differences can account for clinical heterogeneity within a molecularly defined group of leukemias.

[Immunologic and genetic markers of exposure to aromatic hydrocarbons in workers].

Workers exposed to aromatic hydrocarbons appeared to have prevalence of heterozygous variants of CYP1A1 gene (9893 A/G) and tumor necrosis factor gene reliably higher vs. the reference group 2.5 and 3.3 times respectively, and level of anti-benzene antibodies (IgG) increased vs. the reference group. The data presented demonstrate negative immunogenetic associations of aromatic hydrocarbons influence on oil extraction operators.

MLL-AF9 and FLT3 cooperation in acute myelogenous leukemia: development of a model for rapid therapeutic assessment.

Human leukemias harboring chromosomal translocations involving the mixed lineage leukemia (MLL, HRX, ALL-1) gene possess high-level expression, and frequent activating mutations of the receptor tyrosine kinase FLT3. We used a murine bone marrow transplant model to assess cooperation between MLL translocation and FLT3 activation. We demonstrate that MLL-AF9 expression induces acute myelogenous leukemia (AML) in approximately 70 days, whereas the combination of MLL-AF9 and FLT3-ITD does so in less than 30 days. Secondary transplantation of splenic cells from diseased mice established that leukemia stem cells are present at a very high frequency of approximately 1:100 in both diseases. Importantly, prospectively isolated granulocyte macrophage progenitors (GMPs) coinfected with MLL-AF9 and FLT3-ITD give rise to a similar AML, with shorter latency than from GMP transduced with MLL-AF9 alone. Cooperation between MLL-AF9 and FLT3-ITD was further verified by real-time assessment of leukemogenesis using noninvasive bioluminescence imaging. We used this model to demonstrate that MLL-AF9/FLT3-ITD-induced leukemias are sensitive to FLT3 inhibition in a 2-3 week in vivo assay. These data show that activated FLT3 cooperates with MLL-AF9 to accelerate onset of an AML from whole bone marrow as well as a committed hematopoietic progenitor, and provide a new genetically defined model system that should prove useful for rapid assessment of potential therapeutics in vivo.