Modifying the Cyanobacterial Metabolism as a Key to Efficient Biopolymer Production in Photosynthetic Microorganisms.

Cyanobacteria are photoautotrophic bacteria commonly found in the natural environment. Due to the ecological benefits associated with the assimilation of carbon dioxide from the atmosphere and utilization of light energy, they are attractive hosts in a growing number of biotechnological processes. Biopolymer production is arguably one of the most critical areas where the transition from fossil-derived chemistry to renewable chemistry is needed. Cyanobacteria can produce several polymeric compounds with high applicability such as glycogen, polyhydroxyalkanoates, or extracellular polymeric substances. These important biopolymers are synthesized using precursors derived from central carbon metabolism, including the tricarboxylic acid cycle. Due to their unique metabolic properties, i.e., light harvesting and carbon fixation, the molecular and genetic aspects of polymer biosynthesis and their relationship with central carbon metabolism are somehow different from those found in heterotrophic microorganisms. A greater understanding of the processes involved in cyanobacterial metabolism is still required to produce these molecules more efficiently. This review presents the current state of the art in the engineering of cyanobacterial metabolism for the efficient production of these biopolymers.

Adiponectin, leptin and IL-1 ß in elderly diabetic patients with mild cognitive impairment.

The aim of the study was to determine the serum levels of adiponectin, leptin and IL-1 ß in elderly diabetic patients with and without mild cognitive impairment (MCI) and to examine the associations of these markers with clinical and cognitive parameters. A biochemical evaluation was performed of 62 seniors with type 2 diabetes (T2DM) and MCI, and 132 seniors with T2DM but without MCI (controls). Serum leptin and IL-1 ß levels were higher and adiponectin concentration was lower in MCI patients than controls. In MCI subjects, adiponectin level was negatively correlated with leptin, IL-1 ß levels and BMI. Leptin concentration was correlated with IL-1 ß level. Univariate logistic regression models revealed that the factors which increased the likelihood of diagnosis of MCI in elderly patients with T2DM were higher levels of HbA1c, leptin, IL-1 ß and triglycerides, as well as lower levels of adiponectin and HDL cholesterol. Similarly, previous CVD, hypertension, hyperlipidemia, retinopathy, nephropathy, hypoglycemia, longer duration of diabetes, increased number of co-morbidities, older age, fewer years of formal education were found to be associated with MCI. The multivariable model indicated fewer years of formal education, previous CVD, hypertension, increased number of co-morbidities, higher HbA1c and IL-1 ß levels and lower adiponectin level. Elderly diabetic patients with MCI have higher levels of leptin and IL-1 ß and lower levels of adiponectin. Further prospective studies are needed to determine the role of these markers in the progression to dementia.

Plasma levels of thrombomodulin, plasminogen activator inhibitor-1 and fibrinogen in elderly, diabetic patients with depressive symptoms.

BACKGROUND: Diabetes, depression and aging have been associated with pro-inflammatory and prothrombotic state. AIM: The aim of the study was to determine the plasma levels of thrombomodulin, plasminogen activator inhibitor-1 (PAI-1) and fibrinogen in elderly diabetic patients with and without depressive symptoms and to examine factors (including thrombomodulin, PAI-1, fibrinogen levels) associated with depressive symptoms in elderly patients with type 2 diabetes (T2DM). METHODS: A total of 276 T2DM elders were evaluated: 82 subjects with depressive symptoms and 194 controls. Data were collected concerning biochemical parameters and biomarkers. RESULTS: Plasma thrombomodulin, PAI-1 and fibrinogen were elevated in patients with depressive symptoms compared to controls. Thrombomodulin level was correlated with fibrinogen and PAI-1 levels. All parameters were correlated with the Geriatric Depression Scale-30 score. The univariate logistic regression models revealed that variables which increased the likelihood of diagnosis of depressive symptoms in elderly patients with T2DM were: female sex, smoking habit, longer duration of T2DM, hyperlipidemia, neuropathy, increased number of co-morbidities, higher BMI, and higher levels of total and LDL cholesterol, thrombomodulin, PAI-1 and fibrinogen. In addition, the multivariable analysis indicated that female sex, smoking habit, increased number of co-morbidities, higher BMI, and higher levels of LDL cholesterol and thrombomodulin are the predisposing factors for depressive symptoms. CONCLUSIONS: Elderly diabetic patients with depressive symptoms have higher levels of thrombomodulin, PAI-1 and fibrinogen. Further prospective larger studies are needed to provide potential directions for the research, treatment and prevention of co-morbid depression and diabetes.

Short-term reproducibility of the inflammatory phenotype in different subgroups of adult asthma cohort.

Inflammatory phenotype classification using induced sputum appears attractive as it can be applied to inflammation-based management of the patients with asthma. The aim of the study was to determine the reproducibility of inflammatory phenotype over time in patients with asthma. In 66 adults asthma was categorized as steroid-naïve (SN, n = 17), mild to moderate (MMA, n = 33), and refractory treated with oral corticosteroids (RA, n = 16). Clinical assessment, skin prick testing, spirometry, and two sputum inductions in 4-6-week interval were done. Inflammatory phenotypes were classified as eosinophilic (EA), consisting of eosinophilic and mixed granulocytic phenotypes, and noneosinophilic (NEA) consisting of paucigranulocytic and neutrophilic phenotypes. During study asthma treatment remained constant. In SN group 25% of patients changed phenotype from EA to NEA and 44% changed phenotype from NEA to EA. In MMA group 26% of patients changed phenotype from EA to NEA and 50% changed phenotype from NEA to EA. In 29% of RA patients inflammatory phenotype changed from EA to NEA and in 22% it changed from NEA to EA. Inflammatory classification, using induced sputum, is not fully reproducible in adults with asthma in short-term evaluation. EA seems to be more stable phenotype across all subgroups whereas NEA remained stable only in RA group.

Evaluation of Markers of Inflammation and Oxidative Stress in COPD Patients with or without Cardiovascular Comorbidities.

BACKGROUND: Although both chronic obstructive pulmonary disease (COPD) and cardiovascular diseases (CVD) are characterised by chronic, systemic inflammation, their reciprocal interactions are poorly understood. The purpose of this study was to determine the concentrations of both inflammatory and oxidative stress biomarkers in the serum and exhaled breath condensate (EBC) of COPD patients, either with coexisting CVD or without cardio-vascular comorbidities. METHODS: Twenty-four COPD patients with CVD were allocated to group A, 20 COPD patients without CVD were assigned to group B and 16 healthy patients were included as a control. A medical history and physical examination were performed, and the following were measured: serum CRP concentration, glucose level, uraemic acid level and lipid profile. In addition 8-isoprostane, LTB4 and IL-8 concentrations were measured both in serum and EBC. Spirometry, six-minute walk test and echocardiography were performed in all subjects. RESULTS: EBC concentrations of 8-isoprostane and LTB4, and serum levels of CRP, 8-soprostane, LTB4, IL-8 were significantly higher in COPD patients than in healthy controls. COPD patients with CVD were not found to have higher concentrations of the assessed markers than those without CVD, neither in the serum nor EBC. CRP, 8-isoprostane and LTB4 levels in serum, and IL-8 concentration in EBC correlated negatively with the value of forced expiratory volume in one second. CONCLUSIONS: Although systemic inflammation coexists with COPD, it is not elevated in COPD patients with CVD. Since this phenomenon may result from treatment with statins, future studies should state whether COPD patients could benefit from the additional statin therapy.

Pneumococcal and seasonal influenza vaccination among elderly patients with diabetes.

Both seasonal influenza vaccination and pneumococcal vaccination are recommended for elderly diabetics. The aim of the study was to determine the rate of seasonal influenza vaccination over the previous twelve months, pneumococcal vaccination over a lifetime, and to identify predictors which affect likelihood of vaccination. 219 diabetics elders were detailed questioned 3 months after the end of 2012/2013 influenza season. 26.48% of patients have been vaccinated against influenza in the last year and only 9.13% of patients reported pneumococcal vaccination in the past. The logistic regression analysis revealed that variables which increased the likelihood of having been vaccinated against influenza were: higher number of anti-hyperglycemic medications, increased number of co-morbidities, higher patients' income, recommendation of vaccination from General Practitioners (GPs) and specialist. Significant predictors of pneumococcal vaccine uptake included increased number of co-morbidities and recommendation of vaccination received from GPs and specialist. The commonest reasons given by those unvaccinated were lack of information about immunization and low perceived benefits of vaccination. Of patients who were not treated with influenza vaccine 86.7% had never received recommendation from specialist and 71.4% had never been advised by GPs. Influenza vaccination was too expensive to 24.85% of patients. The vaccination rate among elderly diabetics in Poland is low. Lack of knowledge and patients' income are the main barriers. Increased awareness of healthcare professionals to educate and encourage vaccination and propagation of free vaccinations to all people at risk may increase the rate of vaccination against influenza and pneumococcal disease.

Concentration of angiopoietins 1 and 2 and their receptor Tie-2 in peripheral blood in patients with chronic obstructive pulmonary disease.

INTRODUCTION: Both angiopoietins (angiopoietin 1 - Ang-1, angiopoietin 2 - Ang-2) and angiopoietin receptors (Tie) are involved in angiogenesis and vascular remodeling. AIM: To assess concentrations of Ang-1, Ang-2 and Tie-2 in blood of patients with chronic obstructive pulmonary disease (COPD) and evaluate if their concentrations depend on the severity of the disease. MATERIAL AND METHODS: Thirty patients with COPD (stage II-IV) and 8 healthy smokers as well as 8 healthy non-smokers were included in the study. Detailed history was taken, physical examination and spirometry tests were done and blood samples were taken for evaluation of serum concentrations of Ang-1, Ang-2 and Tie. RESULTS: Among COPD patients, 8 patients suffered from moderate disease, 8 patients had severe, while 14 patients had very severe disease. The concentrations of Ang-1 and Ang-2 were not significantly greater in patients with COPD than in healthy controls. The highest concentrations of Ang-1 and Ang-2 were observed in patients with moderate COPD, and levels of Ang-2 correlated with Tie-2 in this group of patients. The levels of Ang-1 were the lowest in healthy non-smokers and in patients with severe COPD, where they inversely correlated with Tie-2. The concentrations of Ang-2 were not significantly higher in patients with moderate COPD when compared with those with severe and very severe disease and healthy smokers, and were significantly higher than in healthy non-smokers. CONCLUSIONS: It is possible that Ang-1, Ang-2 and Tie-2 play an important role especially in the early stage of COPD but not in the late phase when vascular complications of the disease occur.

Wheal and flare reactions in skin prick tests of patients treated with montelukast alone or in combination with antihistamines.

BACKGROUND: Because antileukotrienes may inhibit inflammation, it is plausible that montelukast administered for a long time could suppress skin wheal and flare reaction, and thus, it should be discarded prior to the tests. This study assessed the effect of long-lasting treatment with montelukast alone or in combination with antihistamines on wheal and flare in skin pricks tests (SPT) in patients sensitized to perennial allergens. METHODS: We conducted a 32-week, double-blind, placebo-controlled, cross-over and randomized trial that implicated two arms: arm A, 20 patients received levocetirizine, montelukast with or without levocetirizine or placebo; arm B, 20 patients received desloratadine, montelukast with or without desloratadine or placebo. All treatment periods lasted 6 weeks and were separated by 2-week washouts. At baseline and on the last day of each treatment period, SPT were performed in all participants. RESULTS: Both levocetirizine and desloratadine in monotherapy, or in combination with montelukast, were effective in reducing wheal and flare in SPT. Monotherapy with montelukast did not change the size of the wheal for either histamine or for house dust mites, in either arm of the study, but significantly reduced the size of flare for histamine in arm A. Addition of montelukast to antihistamine did not exceed efficacy of monotherapy with antihistamine in both arms of the study. CONCLUSIONS: Since the size of wheal determines the results of SPT, montelukast, even taken for a long time, does not have to be discarded prior to the tests.

Risk factors associated with irreversible airway obstruction in nonsmoking adult patients with severe asthma.

Persistent airways obstruction (PAO) may affect some patients with severe asthma and may significantly worsen the prognosis. This study was designed to detect risk factors associated with persistent airflow limitation in nonsmoking adult patients with severe asthma. A total of 68 adults with severe asthma were recruited and followed prospectively for four to six weeks during the stable phase of disease. For all patients, at every visit spirometry with reversibility test was performed. Based on the results, patients were stratified into group 1 (reversible obstruction) or group 2 (PAO). In both cohorts, associations of postbronchodilatator forced expiratory volume in one second/forced vital capacity ratio (FEV1/FVC) with patients' age, gender, asthma duration, history of atopy and allergy, family history, medications, frequency of previous exacerbations, infections, hospitalizations, and artificial ventilation due to the asthma attack-related respiratory failure were investigated. Using a univariate logistic regression analysis, we have shown that older age, more than six exacerbations per year, artificial ventilation in the past, at least one hospitalization per year, the presence of atopic dermatitis, and exposure to domestic visible mold were all independent risk factors of PAO. Furthermore, multivariate regression analysis demonstrated that especially those with domestic exposure to visible molds, with very frequent exacerbations and with at least one hospitalization throughout the last year, were at risk for developing PAO. Domestic exposure to molds, hospitalization during the last year, and very frequent exacerbations were associated with PAO in patients with severe asthma. These factors may help in predicting fixed airflow limitation in nonsmoking patients with severe asthma.

Switching from systemic steroids to ciclesonide restores the hypothalamic pituitary-adrenal axis.

INTRODUCTION: Treatment of difficult asthma with oral corticosteroids (OCS) may suppress the hypothalamic-pituitary-adrenal axis. AIM: In this study we have checked if the substitution of OCS with very high doses of ciclesonide may restore the adrenal function without losing the control of the disease. MATERIAL AND METHODS: In 5 patients with difficult, uncontrolled asthma despite treatment with OCS, inhaled and systemic glucocorticosteroids were replaced with very high doses of ciclesonide (1600-2400 µg/day). The symptoms of asthma and the lung function were assessed at baseline and on the 28(th), 56(th) and 70(th) day of treatment, whereas the levels of cortisol and adrenocorticotropic hormone (ACTH) in the morning were measured at baseline and on the 28(th) and the 56(th) day of treatment. RESULTS: In all patients, the control of asthma symptoms, measured with Asthma Control Test questionnaire, improved from the mean score of 9.4 to 19.8 in 70 days. In 4 subjects force expiratory volume in 1 s improved gradually through the entire study reaching a mean improvement of 585 ml in 70 days. The ACTH levels were normalized in 3 patients after 28 days of observation and in all patients after 56 days. The cortisol level was normalized in 4 patients after 28 days and in another subject after 56 days of treatment with ciclesonide. CONCLUSIONS: Switching from prednisone to very high doses of ciclesonide normalized the hypothalamic-pituitary adrenal axis function and also improved the disease control and the lung function in these 5 patients with difficult asthma.

Association between Serum Irisin and Leptin Levels and Risk of Depressive Symptoms in the Diabetic Elderly Population.

BACKGROUND: Adipokines are considered to be involved in the pathogenesis of diabetes and depression. The associations of serum levels of leptin and irisin with depressive symptoms were investigated in elderly patients with type 2 diabetes (T2DM). METHODS: 189 elderly diabetics were assessed with the 30-item Geriatric Depression Scale (GDS-30), and 57 patients with depressive symptoms and 132 controls were selected. Blood biochemical parameters, including serum irisin and leptin, were measured. RESULTS: Serum irisin levels were decreased and leptin concentrations were significantly higher in T2DM patients with depressive symptoms compared to controls. In all subjects, the irisin level was inversely correlated with the leptin level and the GDS-30 score, whereas the leptin level was highly correlated with BMI and the GDS-30 score. Higher levels of leptin and lower concentrations of irisin are, among other factors, variables indicative of predictive capacity for depressive symptoms in elderly patients with T2DM. CONCLUSIONS: The results indicated that irisin and leptin levels may be used as diagnostic markers of depressive symptoms in diabetic, elderly patients and as potential therapeutic targets for the treatment. Further prospective and more extensive studies are needed to clarify the role of these adipokines in the common pathogenesis of depression and diabetes.

sICAM-1 and TNF-α in asthma and rhinitis: relationship with the presence of atopy.

BACKGROUND: A genetically determined overproduction of specific immunoglobulin E (IgE) underlies many diseases like asthma or allergic rhinitis. IgE as well as tumor necrosis factor-α (TNF-α), and intercellular adhesion molecule-1 (ICAM-1) play a critical role in the induction and maintenance of inflammation. While the correlation between IgE and atopy is inseparable, little is known about the correlation of atopy with markers of inflammation. OBJECTIVE: We investigated the relationship between the serum concentrations of TNF-α, soluble ICAM-1 (sICAM-1), and the presence of atopy in patients with persistent rhinitis or asthma. METHODS: Serum concentrations of sICAM-1, TNF-α, and total IgE were investigated in 64 adults with persistent allergic rhinitis, 17 subjects with nonatopic rhinitis, 90 patients with asthma, and 21 healthy individuals. Atopy was diagnosed on the basis of positive family history, skin prick tests, and serum IgE concentration. RESULTS: Total IgE concentration was significantly higher in patients with atopic rhinitis or asthma when compared with nonatopic patients and healthy individuals and was the highest in patients suffering from severe atopic asthma who were not treated with systemic glucocorticosteroids. Although there were marked alterations in IgE in atopic and nonatopic patients, there were no significant differences between atopic and corresponding groups of nonatopic rhinitic and asthmatic patients in sICAM-1 and TNF-α concentrations. (sICAM-1 in rhinitis: atopic vs. nonatopic patients: 224.02 and 221.08 ng/ml, respectively, p > .05; in mild/moderate asthma: atopic vs. nonatopic: 306.22 and 326.39 ng/ml, respectively, p > .05; severe asthma without oral corticosteroids therapy: atopic vs. nonatopic: 418.03 and 468.09 ng/ml, respectively, p > .05; and severe asthma with oral corticosteroids therapy: atopic vs. nonatopic: 320.66 and 308.09 ng/ml, respectively, p > .05). CONCLUSIONS: Concentrations of sICAM-1 and TNF-α are significantly higher in patients with asthma compared with those observed in patients with rhinitis, but they are independent of the presence of atopy.

Intravenous immunoglobulin replacement therapy in the treatment of patients with common variable immunodeficiency disease: an open-label prospective study.

BACKGROUND: Common variable immunodeficiency (CVID) is characterized by humoral immunodeficiency resulting in increased susceptibility to infections and diminished responses to protein and polysaccharide vaccines. Intravenous immunoglobulins (IVIgs) constitute a replacement therapeutic regimen for CVID and other primary and selected secondary immunodeficiencies but their mode of action is still not fully understood. OBJECTIVE: The purpose of this study was to assess the effect of IVIg replacement therapy on the population of regulatory T cells (cells expressing CD4, CD25 and low levels of CD127) [T(regs)]), plasma levels of interleukin (IL)-2 and IL-10, and expression of fragment, crystallizable γ receptor IIb (Fc γ RIIb) [CD32b] on CD19+ B cells in CVID patients. METHODS: This was an open-label prospective trial that included 17 CVID patients and seven healthy subjects as case controls. The diagnosis of CVID was primarily established by clinical criteria designed by the European Society for Immunodeficiencies (ESID) and was confirmed by low serum levels of two out of three subclasses of immunoglobulins (IgG, IgA or IgM). All CVID patients were treated with the IVIg preparation Flebogamma® 5%, a highly purified, pasteurized normal human IgG extracted from the serum of healthy individuals, administered at a dose of 300 mg/kg by slow 2-hour intravenous infusion. Blood samples were collected 30 minutes before the infusion and 30 minutes and 2 weeks after the termination of the infusion. We examined: (i) the plasma levels of IL-2 and IL-10; (ii) the percentage of CD4+ T cells and T(regs); and (iii) the expression of Fc γ RIIb on the surface of CD19+ B cells. RESULTS: CVID patients had higher plasma levels of IL-2 (p = 0.045) and IL-10 (p = 0.002) as well as a higher expression of Fc γ RIIb on CD19+ B cells (p = 0.0119) before IVIg compared with healthy controls. The infusion of IVIg led to further increases in the plasma levels of these cytokines 30 minutes after the termination of the infusion versus baseline (IL-2: p = 0.0004; IL-10: p = 0.0003). IVIg did not affect the expression of Fc γ RIIb. Finally, IVIg infusion resulted in elevation of the percentages of CD4+ T cells (p = 0.028) and T(regs) (p = 0.006) in the blood 30 minutes after the infusion. CONCLUSION: Flebogamma® 5% as replacement therapy not only supplies immunoglobulins but also modulates the immune response, and in this way may provide additional benefits to patients with CVID.

Exhaled 8-isoprostane in sarcoidosis: relation to superoxide anion production by bronchoalveolar lavage cells.

OBJECTIVE: This study was designed to examine the mutual relationship between 8-isoprostane in exhaled breath condensate (EBC) and superoxide anion generation by bronchoalveolar lavage fluid (BALF) cells in patients with sarcoidosis. DESIGN: About 29 patients with sarcoidosis, 34 healthy never smokers (control group for EBC) and 15 healthy never smokers (control group for BAL) were examined. EBC was collected directly before bronchoscopy. 8-Isoprostane was measured by ELISA, and superoxide anion by colorimetry. RESULTS: Exhaled breath condensate 8-isoprostane is increased in sarcoidosis (median, 25-75 percentile): 2.50; 2.50-3.90 versus 6.20; 2.50-16.95 pg/ml, p ≤ 0.05). Spontaneous superoxide anion release from BALF cells was significantly elevated only in patients with a high percentage of lymphocytes in BALF (6.42 ± 1.24 vs. 23.52 ± 4.30 nmol/10(6) cells, p ≤ 0.01). There were no correlations between 8-isoprostane and spontaneous or stimulated superoxide anion release. CONCLUSIONS: We confirmed higher concentrations of EBC 8-isoprostane in sarcoidosis and higher spontaneous release of superoxide anion from BALF cells in patients with sarcoidosis. The increase of EBC 8-isoprostane is not directly related to superoxide anion released from BALF cells.

[Mechanism of action of immunoglobulin applied intravenously]. / Mechanizm dzialania immunoglobulin podawanych dozylnie.

Intravenously applied immunoglobulin (IVIg) are used in medicine over 50 years. Beneficial effects of IVIg therapy have been described in the treatment of primary immunodeficiencies, inflammatory and autoimmune diseases, and new applications are still searched. Nevertheless, the mechanisms responsible for the therapeutic effectiveness of antibodies have not been exactly known. The efficacy of IVIg preparates used in therapeutic doses in infectious and autoimmune diseases indicates on the anti-inflammatory activity resulting from the inhibition of immune cells via different ways. These mechanisms include blocking Fc gamma receptors, modulation of cytokines synthesis and secretion and modulation of antigens on surface of immunocompetent cells. By contrast, IVIg therapy used in the replacement dose gives quite the opposite effect. Substitutive immunoglobulins not only complement the deficiency of antibodies in primary and secondary immunodeficiencies but also have the immunomodulatory and even proinflammatory effect.

Association of hsCRP and vitamin D levels with mild cognitive impairment in elderly type 2 diabetic patients.

AIMS: The aim of the study was to determine the serum levels of 25-hydroxyvitamin D and high-sensitivity C-reactive protein (hsCRP) in elderly diabetic patients with and without mild cognitive impairment (MCI) and to examine factors (including 25-hydroxyvitamin D and hsCRP) associated with MCI in elderly patients with type 2 diabetes (T2DM). METHODS: A total of 194 T2DM elders were evaluated: 62 subjects with MCI and 132 controls. Data was collected concerning biochemical parameters and biomarkers. RESULTS: HsCRP concentration was elevated and 25-hydroxyvitamin D level was decreased in MCI patients to controls. HsCRP level was negatively correlated with 25-hydroxyvitamin D level and with MoCA score, and highly correlated with HbA1c level. The multivariable analysis indicated that less years of formal education, previous CVD and hypertension, increased number of co-morbidities, higher level of hsCRP and lower level of 25-hydroxyvitamin D, are the predisposing factors for MCI. CONCLUSIONS: Higher hsCRP level and lower 25-hydroxyvitamin D may be regarded as a state of cognitive impairment in elderly patients with T2DM. Further prospective larger studies should be conducted to check the association between decreased vitamin D and risk of cognitive decline and to clarify whether this association may be mediated by systemic inflammation.

Improved insulin injection technique, treatment satisfaction and glycemic control: Results from a large cohort education study.

OBJECTIVE: The aim of this study was to elucidate injection techniques, treatment satisfaction and glycemic control after education among patients with type 2 diabetes. METHODS: 4513 insulin-treated diabetic patients enrolled in the observational study EGIDA II (Education and GensuPen In Diabetology II) filled out the questionnaire which focused on key insulin injection parameters, pain sensation scale and satisfaction of the treatment form before (visit 1) and after 3 months treatment with insulin injection (visit 2). The education was performed by trained healthcare professionals. To assess the utility and comfort during using new automatic injection system (GensuPen) we separated 2 groups: A - treated with GensuPen and B - treated with other pens. RESULTS: The education resulted in increased number of patients who properly remix cloudy insulin; inject insulin into skin; change every time the injection site; use the pen needle only once; prepare a pen for injection and store insulin. We noticed significant decrease in BMI and sensation of pain in both groups. Our study revealed that patients' satisfaction with the treatment increased with each of the 5 items (type of the treatment, mood, physical activity, vital energy, a sense of control over the disease) using a 5-point scale, with greater increase in group A. The utility and comfort (weight, thickness, easiness in remove pen cap, cleaning, twisting, keeping in hand the pen, dial the dose, readable signaling of injected dose) during using the GensuPen significantly increased in group A. Finally the mean glucose level in self-control diary was significantly lower after 3 months of the treatment in both groups, however the difference between visit 1 and 2 was greater in group A. CONCLUSION: The study showed that proper selection of pen and professional education can result in the improvement of insulin injection technique, higher patients' satisfaction and better glycemic control.

Monocyte to large platelet ratio as a diagnostic tool for pulmonary embolism in patients with acute exacerbation of chronic obstructive pulmonary disease.

INTRODUCTION    A higher prevalence of pulmonary embolism (PE) has been noted among patients with chronic obstructive pulmonary disease (COPD), particularly in those with acute exacerbations of COPD (AECOPD). Due to a similar clinical presentation and the lack of highly specific laboratory tests, there is a common overuse of computed tomography pulmonary angiography (CTPA). The introduction of an additional, simple, and inexpensive diagnostic tool to help in the diagnosis of PE in patients with AECOPD would be of special interest for everyday clinical practice. OBJECTIVES    The aim of the study was to assess the usefulness of the monocyte to large platelet ratio (MLPR) as a diagnostic tool for PE in patients with AECOPD.  PATIENTS AND METHODS    We performed a retrospective evaluation of patients with AECOPD and suspicion of PE who underwent CTPA. The MLPR was investigated as a marker of thrombosis. Receiver operating characteristics (ROC) curve analyses were preformed to measure the accuracy of the MLPR in comparison with CTPA results and to identify the cutoff value for the MLPR. RESULTS    A total of 101 patients (56 men and 45 women; median age, 72 years; range, 37-94 years) were included in the study. The MLPR showed an excellent accuracy in comparison with CTPA results: the area under the ROC curve was 0.945 (95% confidence interval [CI], 0.904-0.986). The MLPR was characterized by a good accuracy of qualitative test parameters, with high sensitivity (100%; 95% CI, 79.6-100) and specificity (85.7%; 95% CI, 75.9-92.6). CONCLUSIONS    The MLPR measurement appears to be a reliable, simple, inexpensive, and widely available test that may help in the differential diagnosis of PE in patients with AECOPD.

PD-L1 expression and its effect on clinical outcomes of EGFR-mutant NSCLC patients treated with EGFR-TKIs.

OBJECTIVE: Epidermal growth factor receptor (EGFR) activation was reported to upregulate programmed death-ligand 1 (PD-L1) expression in lung cancer cells and subsequently contribute to immune escape, indicating its critical role in EGFR-driven lung tumors. This study characterized PD-L1 expression in patients with surgically resected EGFR-mutant non-small cell lung cancer (NSCLC). The effect of PD-L1 expression on clinical outcomes was also investigated in advanced EGFR-mutant NSCLC treated with EGFR-tyrosine kinase inhibitors (TKIs). METHODS: In total, 73 patients with surgically resected NSCLC and EGFR mutations were identified. PD-L1 expression and CD8+ tumor-infiltrating lymphocyte (TIL) density were assessed by immunohistochemistry. A literature review of publications that assessed the predictive and prognostic value of PD-L1 expression in advanced EGFR-mutant NSCLC patients treated with EGFR-TKIs was performed. RESULTS: Nineteen (26.0%) patients were positive for PD-L1 expression, which was significantly associated with concomitant KRAS mutation (P = 0.020) and marginally associated with higher CD8+ TILs density (P = 0.056). Positive PD-L1 expression was associated with markedly inferior overall survival (OS) in multivariate analysis (P = 0.032). The combination of PD-L1 and CD8+ TILs expression could be used to stratify the population into three groups with distinct prognoses. A meta-analysis of six publications showed that positive PD-L1 expression was not associated with OS [hazard ratio (HR) = 0.90; 95% confidence interval (CI), 0.42-1.38] or progression-free survival (HR = 1.03; 95 CI, 0.73-1.33) in advanced EGFR-mutant NSCLC patients receiving EGFR-TKIs. CONCLUSIONS: PD-L1 expression tended to correlate with CD8+ TIL expression, concomitant KRAS mutation, and poor survival in surgically resected EGFR-mutant NSCLC. PD-L1 expression was neither the predictive nor the prognostic factor in advanced EGFR-mutant NSCLC patients treated with EGFR-TKIs.