Upregulation of Treg-Related Genes in Addition with IL6 Showed the Significant Role for the Distant Metastasis in Colorectal Cancer.

T helper 17 (Th17) and T regulatory (Treg) cytokines appear to be contributing greatly to colorectal cancer (CRC) development and progression. The aim of the current study was to investigate the expression of Foxp3; IL10; TGFB1; IL17A; IL6 and NOS2 genes in tumor tissue, regional positive lymph nodes and distant metastasis obtained from 26 patients with advanced CRC. Quantitative real-time polymerase chain reaction (qPCR) was performed for mRNA detection by TaqMan gene expression assay. In distant metastasis, IL6 was strongly expressed, over 7.5 fold, followed by Treg-related genes Foxp3; IL10 and TGFB1 in contrast to IL17A and NOS2. The similar pattern of expression was observed in positive regional lymph node in addition to significant down-regulation of NOS2 (RQ = 0.287; p = 0.011) and a trend for the elevation of IL17A. In tumor tissue, Fopx3 was significantly upregulated and Foxp3 mRNA positively correlated with TGFB1 in all investigated tissue types. In tumor tissue, expression of IL17A was correlated with NOS2 (r = 0.68; p = 0.005), while in distant metastasis IL10 was in strong relation with TGFB1 and IL6. In addition, a reverse correlation between IL6 and NOS2 (r = -0.66; p = 0.009), was observed in distant metastasis. The simultaneous expression of given Treg and Th17-related genes found both in the primary tumor and in the regional lymph nodes appears to provide suitable microenvironment sufficient for promoting metastatic growth. The upregulation of Foxp3; IL10, TGFB1 and IL6 might be a transcriptional profile hallmark for colorectal metastases.

Oil-soluble vitamins: illegal use for lip augmentation.

Fillers for lip augmentation have become more and more popular in recent years and seem to be indispensable in the cosmetic market nowadays. A series of six young females is presented who developed massive swellings and pain after vitamins A and/or E lip augmentation. The vitamins were extracted from gelatinous capsules (Gericaps [Adipharm EAD, Sofia, Bulgaria], Geritamins [Actavis EAD, Balkanpharma-Dubnitsa AD, Bulgaria], or vitamin E yellow gel capsules) and injected by unprofessional physicians and beauticians in different cosmetic centers. Physical examination revealed firm indurations of the lips and perioral skin, tenderness, erythema, and hard dermal nodules. Histological analysis revealed numerous round-to-ovoid cavities of varying sizes, resulting in a Swiss cheese-like appearance, consistent with lipogranulomas. The patients were treated with systemic and intralesional triamcinolone injections and broad-spectrum antibiotics with good clinical response. In conclusion, these cases demonstrate the danger of the use of unregistered products as fillers injected by unprofessional physicians and beauticians.

MBL2 polymorphisms - manifestations in Bulgarian patients with adult dermatomyositis and systemic lupus erythematosus.

Deficiency in some complement factors is known to cause both systemic lupus erythematosus (SLE) and dermatomyositis (DM). Mannose-binding lectin (MBL) is a recognition molecule of the lectin pathway, and its low levels are reported to influence some autoimmune diseases. Furthermore, MBL2 polymorphisms have been described associated with low MBL serum levels due to impaired MBL structure and function. This is a pilot study to investigate the role of MBL2-550G/C (H/L), -221G/C (Y/X), Arg52Cys (D), Gly54Asp (B), Gly57Glu (C) polymorphisms and MBL serum levels as a risk factor for a development of adult DM and SLE in Bulgarian patients. None of the studied MBL2 polymorphisms appeared associated with the diseases investigated. However, we have found an increased OR of MBL2-221XY genotype in the patients with SLE (OR 1.64, 95%CI 0.77-3.52). MBL2 polymorphisms seemed to affect MBL serum levels and to be associated with the clinical features although none of the associations remained statistically significant after Bonferroni correction. The-550L allele showed an association with electromyography findings in patients with DM. The-221XY genotype was associated with photosensitivity in patients with SLE. The 54AB genotype showed an association with malar rash in patients with SLE, but it appeared decreased among SLE patients with ANA. In conclusion, our results suggest that the MBL2 polymorphisms have rather a disease modifying role and they are not associated with the disease susceptibility in adult DM and SLE among Bulgarian patients.

Sugar ratios, glutathione redox status and phenols in the resurrection species Haberlea rhodopensis and the closely related non-resurrection species Chirita eberhardtii.

Because of their unique tolerance to desiccation, the so-called resurrection plants can be considered as excellent models for extensive research on plant reactions to environmental stresses. The vegetative tissues of these species are able to withstand long dry periods and to recover very rapidly upon re-watering. This study follows the dynamics of key components involved in leaf tissue antioxidant systems under desiccation in the resurrection plant Haberlea rhodopensis and the related non-resurrection species Chirita eberhardtii. In H. rhodopensis these parameters were also followed during recovery after full drying. A well-defined test system was developed to characterise the different responses of the two species under drought stress. Results show that levels of H2O2 decreased significantly both in H. rhodopensis and C. eberhardtii, but that accumulation of malondialdehyde was much more pronounced in the desiccation-tolerant H. rhodopensis than in the non-resurrection C. eberhardtii. A putative protective role could be attributed to accumulation of total phenols in H. rhodopensis during the late stages of drying. The total glutathione concentration and GSSG/GSH ratio increased upon complete dehydration of H. rhodopensis. Our data on soluble sugars suggest that sugar ratios might be important for plant desiccation tolerance. An array of different adaptations could thus be responsible for the resurrection phenotype of H. rhodopensis.

Correlation between TNF-alpha and IL-12p40-containing cytokines in silicosis.

The aim of the present study was to investigate the correlation between tumor necrosis factor-alpha (TNF-alpha) and interleukin (IL)-12p40-containing cytokines, in silicosis patients and healthy donors exposed to silica dust, in an attempt to clarify the reason for variety of clinical outcomes between humans with similar exposure history. Serum levels of TNF-alpha, IL-12p40, IL-12p70 and IL-23 in total group of 62 silicosis patients, 24 healthy donors with similar exposure history like patients and 19 healthy donors without exposure were determined by enzyme-linked-immunosorbent assay (ELISA) method. The serum level of TNF-alpha was significantly higher in healthy donors exposed to SiO(2) (22.4 +/- 11.1 pg/mL) in comparison with non-exposed healthy donors (14.8 +/- 8.8 pg/mL; p = 0.022) and similar to that in silicosis patients. In total, group of silicosis patients significantly elevated levels of TNF-alpha (20.9 +/- 12.9 vs. 14.8 +/- 8.8 pg/mL; p = 0.047) and IL-12p40 (94.5 +/- 51.6 pg/mL vs. 68.7 +/- 26.2 pg/mL; p = 0.029) compared to non-exposed healthy donors were observed. In addition, a strong positive correlation between TNF-alpha and IL-23 levels (r = 0.678; p = 0.022) and between TNF-alpha and IL-12p70 levels (r = 0.75; p = 0.0003) was detected in the group of exposed healthy donors, while in the group of silicosis patients, a significant positive correlation was observed only between TNF-alpha and IL-12p40 (r = 0.434; p = 0.00048), in contrast to other IL-12p40 containing cytokines. In conclusion, we could assume that the elevated serum levels of TNF-alpha are associated with exposition to silica particles, while the elevation of both TNF-alpha and IL-12p40 is associated with silicosis development and severity. Additionally, the balance between IL-12p40-containing cytokines may also contribute to the silicosis progression.

Hereditary keratosis palmoplantaris varians of Wachters (keratosis palmoplantaris striata et areata).

Keratosis palmoplantaris varians of Wachters is a rare autosomal dominant disorder. A 17-year-old boy with characteristic clinical and histopathologic findings of 10 years' duration is described. He displayed yellowish patchy and linear hyperkeratoses of the palms and nummular keratoses on the soles that were symmetrical and often confluent, specifically on the pressure points. Because there was no family history of a similar pattern, he probably represented an isolated case in his family. The typical clinical presentation, differential diagnosis, and classification of hereditary palmoplantar keratoses are discussed.

[Role of IL-12P40 and IL-10 in progression of colorectal cancer].

The aim of study was to investigate the association between serum levels of IL-12p40 and IL-10 and progression of the colorectal cancer (CRC). Be observed an association between severity of CRC and serum levels of investigated cytokines. IL-12p40 was in the highest level in stage-I (423.6 +/- 224.7 pg/ml) compared to more severe stages. In the opposite direction were the data for immunosuppressive cytokine, IL-10. Patients with stage-IV had the highest level of IL-10 (36.02 +/- 9.5 pg/ml). On the basis of our results we could conclude that IL-12p40 and IL-10 serum levels were associated with tumour progression. More severe stages of CRC are characterized by low IL-12p40 and high IL-10 serum levels.

Association of polymorphisms in regulatory regions of interleukin-12p40 gene and cytokine serum level with colorectal cancer.

The present study investigates the association between IL12Bpro and +16974A/C in 3' -untranslated region (UTR) polymorphisms of IL12B and IL-12p40 serum level related to development of colorectal cancer (CRC). Our results showed similar distribution of both the investigated polymorphisms among CRC patients and healthy donors, which suggests that these polymorphisms in IL12B were not associated with the development of CRC. However, we found an increased IL-12p40 level in sera from patients compared to healthy donors and the highest level was observed in stage I compared to more advanced stages of CRC. These findings demonstrated that IL-12p40 serum level has an association with the progression of CRC.

IL-12Bpro and GSTP1 polymorphisms in association with silicosis.

To assess the role of IL-12Bpro and GSTP1 polymorphisms on induced interleukin (IL)-12p40 production in respect to silicosis development, we examined their distribution in 63 silicosis patients and 165 healthy donors. Decreased frequencies of IL-12Bpro-2 allele (42% vs 62.5%, P = 0.0042) and genotype 22 (19% vs 40%, P = 0.02) in patients compared with exposed healthy donors suggested their protective role in silicosis susceptibility. The GSTP1 A/G polymorphism was also associated with silicosis susceptibility. The allele G and genotype GG were overrepresented among patients than among healthy men (36% vs 23%, P = 0.013; 14% vs 2%, P(c) = 0.012). Also, patients' peripheral blood mononuclear cell (PBMC) produced higher IL-12p40 than healthy donors' depending on both IL-12Bpro and GSTP1 polymorphisms. In conclusion, homozygosity of high producer IL-12p40 genotype IL-12Bpro-11 and GSTP1-GG had the highest genetic risk of silicosis development in Bulgarian miners.

Interleukin-12B-3'UTR polymorphism in association with IL-12p40 and IL-12p70 serum levels and silicosis severity.

Susceptibility to silicosis and to disease severity is in part genetically determined. In this study, the role of IL-12B-3'UTR polymorphism in susceptibility and severity of silicosis and its influence on IL-12p40 and IL-12p70 serum level were investigated. The quantity of IL-12p40 and IL-12p70 was detected by enzyme-linked immunosorbent assay and the genotype of IL-12B was determined using the polymerase chain reaction-restriction fragment-length polymorphism method. We observed elevated IL-12p40 in contrast to IL-12p70 serum levels in a group of 62 silicosis patients compared with both control groups. In severe silicosis patients, we detected the highest IL-12p40 serum levels (129.1 +/- 67.7 pg mL(-1)); lower in patients with the moderate (94 +/- 41.6 pg mL(-1)), whereas in mild silicosis, the IL-12p40 levels (67 +/- 23.5 pg mL(-1)) was similar to these in healthy donors. According to IL-12B polymorphism, increased serum levels were observed in subjects with AA genotype (103.2 +/- 46.9 pg mL(-1)) compared to silicosis patients with AC genotype (82.7 +/- 38.3 pg mL(-1)). No significant differences of genotype and allele frequencies of the 3'UTR polymorphism were observed between silicosis patients and healthy controls. However, the heterozygous genotype was found approximately five times more frequently in patients with mild and moderate (48% and 52%) silicosis compared to patients with severe silicosis (11%), and that IL-12B polymorphism may contribute to silicosis severity rather than to susceptibility. Our data demonstrated that elevated serum IL-12p40, independently of IL-12p70 levels, is associated with severity of silicosis, and suggested that IL-12p40 profibrotic activity may contribute to silicosis severity.

Taq-I polymorphism in 3'UTR of the IL-12B and association with IL-12p40 production from human PBMC.

A wide array of studies has demonstrated differences in genotype and allele frequencies of cytokine gene polymorphisms depending on ethnicity and race. In this study, the frequency of Taq-I polymorphism in 3' untranslated region of IL-12B was investigated in two Bulgarian ethnic groups-Bulgarians and Turkish minority. No significant differences of genotype and allele frequencies were observed between these groups. Genotype distribution in the total group of Bulgarian citizens was: AA (61%), CA (32%) and CC (7%), and the allele frequency of 16974 A allele was 0.77. We also evaluated whether this polymorphism affects IL-12p40 production from human PBMC after stimulation. We demonstrated that association between genotype and IL-12p40 production by stimulated PBMC depends on the stimuli used. Our results indicated a significantly decreased IL-12 p40 secretion for the following order of genotypes: AA>CA>CC, after stimulation of PBMC with C3-binding glycoprotein (C3bgp) in contrast to lipopolysaccharide, phytohaemagglutinin and pokeweed mitogen.

Dual role of protein kinase C on mitogen-activated protein kinase activation and human keratinocyte proliferation.

Subconfluent normal human keratinocytes exhibit autonomous (autocrine growth factor driven) proliferation and express the specific markers for keratinocyte proliferation K#5 and K#14. In keratinocyte autocrine culture, the exogenously added epidermal growth factor (EGF) has no effect on cell proliferation and mitogen-activated protein kinase (MAPK) activity. PD98059 inhibits MAPK pathway and autocrine keratinocyte proliferation. Staurosporine and Gö6976 strongly inhibit autonomous keratinocyte proliferation. In contrast, Gö6983 (which does not inhibit PKC micro ) inhibits only 20% of autocrine keratinocyte proliferation. Staurosporine inhibits MAPK activity, whereas Gö6976 and Gö6983 strongly increase it. We have concluded that MAPK, PKC micro and probably PKCalpha take part in autocrine keratinocyte proliferation. The effect of Gö6976 and Gö6983 on MAPK activity could be explained by the inhibition of PKC-dependent MAPK-phosphatase expression. The effect of staurosporine could be explained by its paradoxical action (activation) on protein kinase C (PKC) in keratinocytes.

Ca(2+)/calmodulin-dependent protein kinase (CaM-kinase) inhibitor KN-62 suppresses the activity of mitogen-activated protein kinase (MAPK), c-myc activation and human keratinocyte proliferation.

Autocrine growth of human epidermal keratinocytes can be maintained in subconfluent cell cultures in the absence of exogenous growth factors. We used this culture model to investigate the interactions between the mitogen-activated protein kinase (MAPK) pathway and Ca(2+)/calmodulin-dependent protein kinases (CaM-kinases) in autocrine keratinocyte proliferation. We have previously demonstrated that MAPK and protein kinase C (PKC) are both involved in keratinocyte proliferation in a complex set of interactions. Treatment of keratinocytes with PD98059, a potent inhibitor of MAPK kinase, inhibited the MAPK pathway, c-myc activation and autocrine keratinocyte proliferation. Application of the CaM-kinase inhibitor KN-62 also led to a strong inhibition of MAPK/c-myc activation and autocrine keratinocyte proliferation. Other inhibitors, such as wortmannin (selective and potent inhibitor of phosphatidylinositol 3-kinase) and AG 490 (JAK2 inhibitor) had weak effects on autocrine keratinocyte proliferation, MAPK and c-myc activation. Our results clearly demonstrate a crosstalk between CaM-kinase/MAPK pathways in transducing keratinocyte proliferation stimuli.