Chemical Composition and Antibacterial Activity of Liquid and Volatile Phase of Essential Oils against Planktonic and Biofilm-Forming Cells of Pseudomonas aeruginosa.

Pseudomonas aeruginosa is an opportunistic pathogen causing life-threatening, hard-to-heal infections associated with the presence of a biofilm. Essential oils (EOs) are promising agents to combat pseudomonal infections because of the alleged antimicrobial activity of their volatile fractions and liquid forms. Therefore, the purpose of this paper was to evaluate the antibacterial efficacy of both volatile and liquid phases of seven EOs (thyme, tea tree, basil, rosemary, eucalyptus, menthol mint, lavender) against P. aeruginosa biofilm and planktonic cells with the use of a broad spectrum of analytical in vitro methods. According to the study results, the antibacterial activity of EOs in their liquid forms varied from that of the volatile fractions. Overall, liquid and volatile forms of rosemary EO and tea tree EO displayed significant antibiofilm effectiveness. The outcomes indicate that these particular EOs possess the potential to be used in the therapy of P. aeruginosa infections.

CD40 stimulation induces differentiation of acute lymphoblastic leukemia cells into dendritic cells.

UNLABELLED: Despite the very high percentage of long-term remissions in acute lymphoblastic leukemia (ALL) in children, some of them suffer from recurrence of the disease. New treatment modalities, e.g. effective geno- and immunotherapy are needed. The use of neoplasmatic cells to present tumor antigens is one of the approaches in cancer vaccines. ALL cells lack the expression of costimulatory molecules and are poor antigen presenting cells (APCs) for T-cell activation. CD40/40L interaction stimulates B-cells to proliferate, differentiate, upregulate costimulatory molecules and increase antigen presentation. The aim of the study was to test the hypothesis that ALL cells can be turned into professional APCs by CD40L activation. Children with B-cell precursor ALL were enrolled into the study. Mononuclear cells from bone marrow or peripheral blood were stimulated with CD40L and interleukin 4. RESULTS: 1) after culture we noted upregulation of all assessed costimulatory, adhesion and activatory molecules i.e. CD1a, CD11c, CD40, CD54, CD80, CD83, CD86, CD123, HLA class I and II; 2) CD40L activated ALL cells induced proliferation of allogeneic T-cells (measured by [(3)H]thymidine incorporation). These results confirm the possibility of enhancing the immunogenicity of ALL cells with the CD40L system and indicate that this approach can be used in immunotherapeutic trials.

[Nitric oxide in supernatants of leukocytes and the serum of patients with B-CLL]. / Tlenek azotu w nadsaczach hodowli leukocytów i surowicy krwi u pacjentów z przewlekla bialaczka limfatyczna B-komórkowa (pbl-B).

Progression of B-cell chronic lymphocytic leukemia (B-CLL) is linked to an abnormal immune system in the host. Recent studies have suggested that polymorphonuclear neutrophils (PMN) play a role in the malignancy process through release of a wide range of mediators, involving nitric oxide (NO). The aim of this study was to examine NO production by PMN and, for comparison of peripheral blood mononuclear cell (PBMC) confronted with the expression and concentration of inducible NO synthase (iNOS) in these cells derived from patients with B-CLL. Results obtained were analyzed according to Rai' staging systems. Our results have shown impaired production of NO by human neutrophils and mononuclear cells. Furthermore, higher expression of iNOS detected by western blot as well as increased concentrations iNOS estimated by ELISA in these cells were observed. We also found higher expression and concentration of iNOS in PMN and PBMC patients in III stage in comparison with patients in I stage of the disease. Additionally we demonstrated a lower production of superoxide anion by neutrophils of patients with B-CLL. Results obtained suggest that impaired NO production, despite of enhanced expression of iNOS, may have a favorable effect on anti-tumor response in patients with B-cell chronic lymphocytic leukemia.

[Estimation of L-selectin expression on neutrophils and level of soluble L-selectin form in serum of patient with chronic myelogenic leukemia]. / Ocena ekspresji L-selektyny na neutrofilach oraz rozpuszczalnej formy sL-selektyny w surowicy krwi u chorych na przewlekla bialaczke szpikowa.

The aim of this study was the estimation of L-selectin expression on PMN and concentration of sL-selectin in patients serum with chronic myelogenic leukemia. The results indicate the increased expression of L-selectin on isolated neutrophils from peripheral blood of patients with chronic myelogenic leukemia. A concentration of sL-selectin was also increased in patients serum with chronic myelogenic leukemia. High concentration of L-selectin on PMN makes binding neoplastic cells easier. Increased level of sL-selectin might activate of the adhesion process in patients with chronic myelogenic leukemia. High expression of L-selectin on PMN may be a response to higher levels of TNF-alpha in serum blood patients with chronic myelogenic leukemia.

TNF family molecules in the serum of patients with B-cell chronic lymphocytic leukemia (B-CLL).

Tumor necrosis factor (TNF) superfamily, involving membrane receptors and ligands are important for the growth and survival of leukemic B cells. In the present study levels of TNF-alpha, sTNFRp55, sTNFRp75 and sCD40 and sCD40L in the serum of patients with B-CLL before and after treatment were measured. In sera of patients before treatment increased concentrations of sCD40 and decreased concentrations of sCD40L were shown. Increased concentrations of TNF-alpha and sTNFRp75 and lack of changes in sTNFRp55 concentrations were also found. Results obtained suggest that the relationships between examined soluble form of TNF family proteins may influence the development of B-cell chronic lymphocytic leukemia. The used therapy with 2CdA and CMC led to a favorable effect for host through changes in the relations between sCD40 and sCD40L. It was also found that sCD40 and sCD40L serum concentrations, which are dependent on the clinical stage and used therapy, are more sensitive tumor markers than TNF-alpha and its soluble receptor in patients with B-CLL treated with 2CdA and CMC.

[Estimation of level of soluble form PECAM-1, ICAM-2 and TNF-alpha, IL-18 in serum patients with chronic myelogenic leukemia]. / Ocena stezenia rozpuszczalnych form PECAM-1 i ICAM-2 i stezenia TNF-alpha i IL-18 w surowicy krwi pacjentów z przewlekla bialaczka, szpikowa.

The aim of this study was to estimate sPECAM-1, sICAM-2 and TNF-alpha and IL-18 concentrations in serum patients with chronic myelogenic leukemia. The results indicate of increased level sPECAM-1, sICAM-2 and TNF-alpha, IL-18 concentrations in serum patients with chronic myelogenic leukemia. Elevation levels of sPECAM-1 and sICAM-2 may lead to inhibit of making myelogenic leukemia cells infiltrations through the block of surface their receptors in patients with CML. High concentration of TNF-alpha and 11-18 in blood serum may indicate high expression of sPECAM-1 by activated specific enzymes responsible for releasing sPECAM-1.

[Subclinical hyperthyrodism, diagnosis and radioiodine therapy]. / Subkliniczna nadczynnosc tarczycy, diagnostyka i leczenie radioizotopowe.

Subclinical hyperthyroidism is a state of increased thyroid function with few or no clinical definitive signs or symptoms of hyperthyroidism. It is characterised by a decrease of serum (TSH) concentration below 0.1 mU/L, when serum levels of total and free thyroxin and triiodothyronin concentration are within normal reference ranges. It is not a rare finding and rates between 0.02% and 11.3% have been reported in different groups. The clinical diagnosis of subclinical hyperthyroidism is very difficult in the absence of the typical symptoms of hyperthyroidism. Therefore the diagnostic evaluation is important, especially with the use of radioisotope scan. In nuclear medicine department we can confirm the provisional diagnosis by the use of thyroid scan. Recent studies reported the effects of subclinical hyperthyroidism on cardiovascular system, skeletal system, cognitive function, on quality of life and life expectancy especially in the elderly patient. Treatment is indicated in the presence of palpitation, or atrial fibrillation, in postmenopausal osteoporosis in women not on hormone replacement therapy, and in elderly patients in whom surgery is contraindicated. According to the opinions of European clinicians and clinician members of the American Thyroid Association, the majority recommend radical treatment for these patients. Radioiodine therapy is considered to be the treatment of choice in most of the patients with nodular goiter.

Apoptosis: its pathophysiology and monitoring. The role of apoptosis in the radioiodine therapy of hyperthyroidism.

The review aims to give an up to date understanding of the mechanisms of apoptosis (programmed cell death), the methods of detecting apoptosis, in particular with regard to imaging such changes non-invasively. Radioiodine (I-131) is a gamma and beta emitting radionuclide and is commonplace in the treatment of hyperthyroidism. I-131 therapy relies on the destruction of thyroid tissue by beta radiation, and such destruction is proposed to be partly as a result of apoptosis. The review undertakes to explore and provoke research into the mechanisms of thyroid cell destruction by I-131, and whether such changes are able to be detected or monitored. Current knowledge concerning apoptosis in the thyroid gland in diseased states (including cancer) are described. The clinical significance of monitoring and modifying apoptosis are emphasized. Furthermore, overt and late destruction of thyroid tissue following I-131 therapy requires elaboration, and the relevance of detecting and modifying thyroid cell apoptosis following I-131 are questioned.

E-selectin, L-selectin, ICAM-1 and IL-6 concentrations changes in the serum of patients with hyperthyroidism in the early period of radioiodine I-131 therapy.

BACKGROUND: Among cytokines- interleukins: -6 and -8 (IL-6, IL-8) and E-selectin (E-sel.), L-selectin (L-sel.) and intercellular cell adhesion molecule-1 (ICAM-1) are the most important links in the initiation of the inflammatory process. Taking into account that the inflammatory process is the basic stage of effective radioiodine therapy, we tried to compare the behaviour of the initial inflammatory factors in the early period of I-131 therapy (RAI) of hyperthyroidism. The aim of the study was to estimate the behaviour of IL-6, ICAM-1, E-selectin and L-selectin concentrations in the serum of patients with hyperthyroidism before and during I-131 therapy. MATERIAL AND METHODS: The groups of 26 patients with Graves' disease (GD) and 18 patients with toxic nodular goiter (TNG), aged 34-77, were studied. Control group (C) consisted of 10 healthy volunteers. For estimation of thyroid function serum concentrations of TSH, free T4 and free T3 were measured by IRMA or RIA kits (Polatom, Poland). IL-6, ICAM-1, E-selectin and L-selectin serum concentrations were determined using ELISA method by Bender kits (USA). Blood samples for all estimations were taken 10-12 days before and in 6th week after I-131 administration. Treatment dose of radioiodine was calculated, basing on modified equation for absorbed dose. RESULTS: Compared to control, no statistical differences in the levels of E-selectin (C--44.4 +/- 11 ng/ml) and L-selectin (C--842 +/- 168.9 ng/ml) were observed before treatment in the patients with GD (E-sel.--59.8 +/- 19.6 ng/ml; L-sel.--1288.2 +/- 273.5 ng/ml) and with TNG (E-sel.--61.5 +/- 18.4 ng/ml, L-sel.--1247.0 +/- 273.5 ng/ml) as well as in the 6th week after I-131 administration; values in GD group were: E-sel.--57.3 +/- 19.5 ng/ml, L-sel.--1142.4 +/- 193.4 ng/ml; in TNG group: E-sel.--62.1 +/- 20.6 ng/ml, L-sel.--1113.5 +/- 236.3 ng/ml. In comparison to control there was no difference in initial IL-6 levels either in GD or in TNG group, but a statistically important decrease was observed in the 6th week after I-131 administration in GD patients (C--2.07 +/- 0.2 ng/ml v. 1.79 +/- 0.16 ng/ml). ICAM-1 serum concentrations before treatment were elevated compared to control group (C--190.2 +/- 34.7 ng/ml) in both groups (GD--263.6 +/- 24.6 ng/ml, p < 0.05; TNG--251.4 +/- 36.1 ng/ml, p < 0.05). In GD patients a statistically significant increase of ICAM-1 was observed in the 6th week (301.1 +/- 33.2 ng/ml, p < 0.05) of RAI whereas in TNG group there was no statistical difference compared to initial values (249.7 +/- 42.6 ng/ml, N.S.). CONCLUSION: We conclude that ICAM-1 and IL-6 may be important factors in the estimation of the inflammatory processes in the thyroid gland during radioiodine therapy, especially GD disease. E- and L-selectins seem to be not helpful in the monitoring of the thyroid inflammatory changes during the early period of I-131 therapy.

The value of estimating serum aproptotic marker concentrations in monitoring and prognosis of 131I--therapy in Graves' disease. Preliminary report.

BACKGROUND: The effect of radioiodine (131I) in Graves' disease (GD) is probably due to the direct physical destruction of thyrocytes by beta radiation, and by the indirect action through stimulation of apoptosis in these cells. The aim of our study was to investigate the changes in serum concentrations of sFas and sFasL as stimulators of apoptosis, and Bcl-2 as an inhibitor of apoptosis in patients with GD following 131I administration. MATERIAL AND METHODS: The study was performed on 30 patients with GD (29 female and 1 male aged 25-45). All patients were euthyroid (biochemical and clinical) prior to radioiodine therapy. The target absorbed dose ranged between 90 and 160 Gy. We assessed markers of apoptosis and hormone concentrations (fT3, fT4 and TSH) in the following manner: before 131I administration, then two weeks, one month, two, three, four, and five months after 131I administration. RESULTS: After four months, the concentrations of sFas and sFasL rose by 50% and decreased during the next month. Pretherapeutic concentrations of Bcl-2 were elevated, and peaked two weeks after ingestion, showing a gradual decrease with time. We found a significant increase in serum TSH, and a decrease of fT3 and fT4 concentrations by the end of the third month of radioiodine therapy. CONCLUSIONS: Decreases in serum levels of sFas and sFasL and increases of Bcl-2 are regarded as characteristic for GD patients before radioiodine therapy. Radioiodine therapy reverses the ratio of estimated markers after four months. The concentrations of hormones reflect actual thyroid function, whereas concentrations of markers of apoptosis may suggest morphological changes.

[Blood levels of the adhesion molecules ICAM-1 and VCAM-1 in patients with chronic myeloid leukemia during remission and exacerbation]. / Badanie zmian stezen czasteczek adhezyjnych ICAM-1 oraz VCAM-1 w okresie zaostrzenia i remisji u chorych na przewlekla bialaczke szpikowa.

A change in ICAM-1 and VCAM-1 concentrations is most likely to occur during the course of chronic myleoid leukemia (CML). The purpose of our study was to examine concentrations of these adhesion molecules in blood plasma, culture supernatant and isolated, broken granulocytes in 20 patients (45-65 years old) with CML in exacerbation and during the remission of the disease and in 10 healthy control subjects. The concentration assay of substances mentioned above was made using ready immunoenzymatic sets of ELISA type. The examinations were carried out on the cultures of cells stimulated and nonstimulated with mitogen--Neupogen of Roche with a the dose of 1 mu/4 ml of culture. Mitogen was added to activate granulocytes and to induce blastic transformation. A significant increase in of plasma ICAM-1 concentration was found in CML exacerbation and remission. The difference between the concentrations of the adhesion molecules with mitogen stimulated and nonstimulated cultures were observed. A significant increase in ICAM-1 and VCAM-1 concentration could suggest a higher secretory function of granulocytes. The values of ICAM-1 were increased in culture supernatants and broken granulocytes before and after adding mitogen in comparison to control groups. The difference in concentration we observed could be characteristic for leukaemic cells.

[Interleukin-6 and interleukin-12 blood levels in patients with chronic B-cell lymphocytic leukemia]. / Stezenia interleukin 6 i 12 we krwi u chorych na przewlekla bialaczke limfatyczna B-komórkowa.

B-cell chronic lymphatic leukaemia (B-CLL) is characterized by proliferation and accumulation of small B-lymphocytes, which are monoclonal in organ. The changes in IL-6 and IL-12 concentrations usually occur during the course of B-CLL. IL-6 and IL-12 seem to be positive kinetic regulators of stem cells. Therefore the purpose of our study was to examine the changes in concentrations of IL-6 and IL-12 in blood plasma, culture supernatant and isolated and broken lymphocytes from patients with B-CLL. The study was performed in I (n = 12) and III (n = 12) stage of disease according to Rai's classification--20 males and 4 females (aged 45-65) and in 12 healthy volunteers blood donors 35-55 years old. The study was approved by the local ethics committee. The measurement of concentrations of IL-6 was performed using the IL-6 immunoenzyme set of ELISA, R&D Systems Europe (UK) in plasma, culture supernatant and broken leukaemic cells. The results showed a significant increase in IL-6 and IL-12 concentration in blood plasma, culture supernatant and inside of the lymphocytes at I and III stage of B-CLL with regard to control groups. An increase of IL-6 and IL-12 concentrations in blood plasma and culture supernatant may suggest higher secretions by lymphocytes these interleukins during the course of B-CLL. An increase of IL-12 in broken leukaemic cells could be characteristic for the biochemistry of malignant lymphocytes.

[Chemokines and their participation in leukemic cells migration]. / Chemokiny i ich udzial w migracji komórek bialaczkowych.

Impaired migration of leukocytes is characteristic feature of leukaemias. Knowledge of the mechanisms of leukaemia cells migration has expanded greatly in recent years. Leukocytes infiltrates are formed in surrounding tissues due to changes in chemokines and adhesion molecules concentrations. The adhesive interactions of cells with other cells and between cells and with the extracellular matrix are started by activation leukaemic leukocytes by specific chemokines. There are four groups of chemokines receptors: CXC, CC, C and CX3C. Unfortunately pathological processes of cells activation in the curse of leukaemias have not been fully explained yet. The paper presents current opinions about structure and role of some chemokines and their receptors in leukaemic cells migration.

[Effect of cytostatic therapy on IL-1beta and L-selectin concentrations, synthesis, accumulation and excretion in patients with acute myeloid leukaemia]. / Próba oceny wplywu leczenia cytostatykami na procesy syntezy, magazynowania oraz wydzielania interleukiny-1beta oraz selektyny L u chorych na ostra bialaczke szpikowa.

The etiology and pathogenesis of the majority of diseases that make up the acute leukemias is unknown. A change in IL-1beta and L selectin concentrations is most likely to occur in the course of subtype M2 of acute myeloid leukaemia (AML). The purpose of our study was to examine the change in concentrations of these molecules mentioned above in blood plasma, culture supernatant and isolated, broken granulocytes in AML patients in both exacerbation and remission of the disease and in healthy control group. Cytokine concentration assay was performed by means of ready immunoenzymatic sets of ELISA type. The examinations were carried out in leukaemic leukocyte cultures Neupogen--stimulated or nonstimulated. Mitogen was added to activate granulocytes and to provoke blastic transformation. A significant increase in IL-1beta concentration was found in AML--exacerbation and remission of the disease in blood plasma, culture supernatant and isolated, broken granulocytes. In all cases L-selectin concentrations were increased in exacerbation and decrease in remission of AML after typical chemotherapy in comparison to controls. A significant increase between the concentrations of cytokines were observed in cultures Neupogen--stimulated and non-stimulated.

[Study of interleukins and their soluble receptors in B-cell chronic lymphatic leukemia]. / Badania stezen interleukin i ich rozpuszczalnych receptorów w przewleklej bialaczce limfatycznej B-komórkowej.

The purpose of our study was to examine concentrations of IL-1 beta, IL-1 beta R, IL-6 and IL-6R in blood plasma, culture supernatant and isolated broken lymphocytes in 20 patients with CLL, at I and III stage of the disease according to Rai's classification and in 10 healthy control subjects. The studies were carried out on the cultures of cells nonstimulated and stimulated with mitogene. A significant increase in IL-1 beta and IL-6 concentrations were found in all study groups during the course of B-CLL. The values of IL-1 beta R and IL-6R were increased in blood plasma at I and III stage of CLL and decreased in culture supernatants and broken lymphocytes before and after stimulation in comparison to control groups. In all cases studied parameters were higher after stimulation. In conclusion, significant increase of IL-1 beta and IL-6 concentrations during CLL may advocacy of higher synthesis and excretions of interleukins--stimulatores of cell proliferation by leukaemic lymphocytes. Increased IL-1 beta R and IL-6R concentrations in blood plasma during CLL, seems to be one of the mechanisms restricted access of IL-1 beta and IL-6 to their surface receptors. An increase of IL-1 beta and IL-6 concentrations and decrease of IL-1 beta R and IL-6R volues suggest survival of autoregulation mechanisms defended against autocrine excreted interleukins. The volues of concentrations of IL-1 beta and IL-1 beta R positively correlated with progress of disease. Such correlation was not found with respect to concentrations of IL-6 and IL-6R.

[Effect of LFA-1 expression on neutrophils and serum level of soluble ICAM-1 in patients with chronic myelogenous leukemia]. / Ocena ekspresji LFA-1 na neutrofilach oraz stezenia rozpuszczalnej formy sICAM-1 w surowicy krwi u pacjentów chorych na przewlekla bialaczke szpikowa.

The aim of this study was to estimation of LFA-1 expression on PMN and concentration of sICAM-1 in serum patients with chronic myelogenic leukemia. The results indicate on increased expression of LFA-1 on isolated neutrophils from peripheral blood of patients with chronic myelogenic leukemia. A concentration of sICAM was increased in serum patients with chronic myelogenic leukemia either. High expression of adhesion molecules can make easier binding neoplasmatic cells to LFA-1 and facilitate their migration to other tissues (distant metastases). On the other hand, soluble forms of ICAM can block connection neoplasmatic cells to membrane forms of adhesion molecules. Probably, the balance between membrane--bound and soluble forms of adhesion molecules is responsible for malignancy progression.

[sCD44 in peripheral blood cultures of mononuclear cells and in the serum of patients with B-cell chronic lymphocytic leukemia]. / Czasteczka sCD44 w hodowlach komórek jednojadrzastych krwi obwodowej i w surowicy krwi u chorych na przewlekla bialaczke limfatyczna B-komórkowa.

The aim of this study was to estimate the ability of peripheral blood mononuclear cells to release of soluble CD44 standard (sCD44st) compared to concentration in the serum of patients B cell chronic lymphocytic leukemia. The results showed the decrease of secretion of sCD44 by leukaemic cells in studied patients. There was no significant differences between patients with I and III stage according to Rai classification. In the serum of patient group we found higher concentrations of sCD44 in comparison with control group. Lack correlation between the concentrations of sCD44 in the cultures and serum suggest that other than leukaemic cells may be significant source of this molecule in patients with B cell chronic lymphocytic leukemia.

The influence of non-radioactive iodine (127I) on the outcome of radioiodine (131I) therapy in patients with Graves' disease and toxic nodular goitre.

BACKGROUND: The aim of the study was to achieve an effective target dose in the thyroid by increasing the effective half-life (Teff) of (131)I by use of iodide ((127)I) two days after (131)I therapy in patients with hyperthyroidism with low Teff. MATERIAL AND METHODS: The study was carried out in two groups. Group A - 41 patients, and Group B - 14 patients, all the patients were with hyperthyroidism with Teff less than 3 days qualified for (131)I therapy. Only group A patients received 600 µg of iodide a day for 3 days, two days after (131)I therapy. Radioiodine uptake (RAIU) after 24 and 48 hours, thyroid scintiscan and ultrasonography were done before and after 12 months of (131)I therapy. RESULTS: In group A a significant increase was seen in the Teff (5 days on average) resulting in an increase in the energy target dose by 28% and 37%, in patients with Graves' disease (GD) and toxic nodular goitre (TNG), respectively. After one year of therapy 50% of GD and 93% of TNG patients achieved euthyroidism; 28% of GD and 3% of TNG patients were in hypothyroidism. In Group B, all the patients had radioiodine treatment failure and received a second therapeutic dose of (131)I. CONCLUSIONS: Administration of (127)I after (131)I treatment can lead to an increase in its effective half-life. This will also increase the absorbed energy dose in thyroid tissue, thereby improving therapeutic outcome without administration of a higher or second dose of (131)I. This may minimize whole-body exposure to radiation and reduces the cost of treatment.