Multilocular infection caused by hypervirulent Klebsiella pneumoniae.

Hypervirulent strains of Klebsiella pneumoniae (hvKP) can cause atypical multilocular infections in otherwise healthy patients. Diagnosis of infection caused by hvKP is based mainly on clinical findings and laboratory results, including detection of virulence genes. It typically manifests as hepatic abscess with metastatic spread. Treatment is based on surgical intervention in combination with targeted antimicrobial therapy. The occurrence of hvKP infection is relatively common in Asia, and while still rare in Europe, incidence is increasing. The article aims to provide a short overview of the issue and increase awareness of the possible occurrence of hvKP infections.

Adipose tissue collection from a living kidney donor; an experimental model for the research of atherosclerosis.

Thanks to an increased number of living-donor kidney transplants the IKEM transplant program offers the possibility of obtaining adipose tissue for scientific purposes from patients with varying degrees of atherosclerosis. Surgery mainly addresses vascular complications of this disease. On the other hand, surgery may also be the reason for the development and acceleration of atherosclerosis - for instance, acceleration of atherosclerosis in the living kidney donor, particularly if, although meeting internationally recognized donation criteria, the donor actually suffers from metabolic syndrome. The effort to refine the examinations of living kidney donors in terms of eliminating the risk of developing atherosclerosis is a long-term project. The aims are to determine the risk factors for living kidney donors and to prevent long-term complications after donation. The paper gives a detailed description of the technique of adipose tissue collection from a living kidney donor and of the experimental model for the research of atherosclerosis.The project has the potential to increase the safety of living kidney donation and to enhance our present knowledge of atherosclerosis development mechanisms.

Could human cold adaptation decrease the risk of cardiovascular disease?

The impact of repeated exposure to cold and cold adaptation on human cardiovascular health is not fully understood. The aim of the study was to determine the effect of cold adaptation on cardiovascular risk factors, thyroid hormones and the capacity of humans to reset the damaging effect of oxidative stress. Ten well cold-adapted winter swimmers (CA) and 16 non-adapted controls (CON) were enroled in this experiment to test whether cold adaptation could influence the parameters of lipoprotein metabolism, cholesterol efflux capacity (CEC), homocysteine, thyroid hormones, antioxidant defence markers (reduced glutathione (GSH), glutathione peroxidase 1 (GPX1), glutathione reductase (GR), catalase (CAT) and paraoxonase 1 (PON1)) and oxidative stress markers (concentration of conjugated dienes (CD)). A decreased apolipoprotein B/apolipoprotein A1 (ApoB/ApoA1) ratio was found in the CA group (p<0.05), but other lipoprotein parameters, including CEC, did not differ significantly. Plasma homocysteine was lower in CA subjects in comparison with controls (p<0.05). Higher triiodothyronine (T3) values were observed in the CA compared to the CON (p<0.05) group, but TSH and other thyroid hormones did not differ between both groups. CA subjects had lower activity of GPX1 (p<0.05), lower concentrations of CD (p<0.05) and increased activities of PON1 (p<0.001) compared to CON subjects. A trend for decreased activity of CAT (p=0.06) in CA compared to CON groups was also observed, but GSH levels did not differ significantly. Zn concentration was higher in the CA group than in the CON group (p<0.001). Human cold adaptation can influence oxidative stress markers. Trends towards the improvement of cardiovascular risk factors in cold-adapted subjects also indicate the positive effect of cold adaptation on cardio-protective mechanisms.

Cholesterol efflux and macrophage polarization in human adipose tissue.

The pro-inflammatory status of adipose tissue (AT) has been found to be related to reverse cholesterol transport (RCT) from peritoneal macrophages. However, this finding was made in experimental models using induced peritonitis and isolated peritoneal macrophages of animals. This experimental relationship is in agreement with RCT changes in man in two extreme situations, sepsis or cardiovascular complications. Given the above, we sought to test RTC in relationship to macrophage polarization in the visceral AT (VAT) of living kidney donors (LKDs) and the effect of conditioned media obtained from their AT. The influence of ATCM on CE capacity was first assessed in an experiment where standard plasma was used as cholesterol acceptor from [14C] cholesterol labeled THP-1. Conditioned media as a product of LKDs' incubated AT showed no effect on CE. Likewise, we did not find any effect of individual plasma of LKDs on CE when individual plasma of LKDs were used as acceptors. On the other hand, we documented an effect of LKDs' adipose cell size on CE. Our results indicate that the pro-inflammatory status of human AT is not likely induced by disrupted RCT but might be influenced by the metabolic status of LKDs' adipose tissue.

[The coincidence of alcoholic liver cirrhosis and chronic pancreatitis]. / Soubezný výskyt alkoholické jaterní cirhózy a chronické pankreatitidy.

UNLABELLED: INTRODUCTION, STUDY AIM: Health consequences of alcoholism vary and include a wide range of pathological conditions from mental disorders to disorders of various parenchymatous organs. There are many differences between alcohol abusers, from the way they abuse alcohol to the nature of bodily harm. The aim of this prospective study was to map and correlate organ involvement with the type of alcohol abuse and smoking and dietary habits. METHODS: A prospective study analyzing a questionnaire on alcoholism, smoking, diet and lifestyle habits completed by 146 patients with liver cirrhosis and chronic pancreatitis. RESULTS: Of the 80 liver cirrhosis patients with the mean age of47.5 year (23-60 years, SD 8.58), 64 were men and 16 (20%) were women. Of the 60 chronic pancreatitis patients with the mean age of 37.8 year (23-60 years, SD 9.66), 60 were men and 6 (9.1%) were women. The mean daily alcohol intake was 57.7 g in patients with cirrhosis and 63.7 g per day in patients with pancreatitis. The first contact with alcohol before 15th year of age was reported by 25.8% of patients with pancreatitis and 8.8% of patients with cirrhosis. Age under 30 years as the period with the largest intake of alcohol was reported by 43.6% of patients with pancreatitis and 20.3% of patients with cirrhosis. There was a statistically significant difference in the type of alcohol used; patients with pancreatitis predominantly drank beer. 16.7% of patients with pancreatitis and 3.7% of patients with cirrhosis started to smoke before 15th year of age. 66.7% of patients with pancreatitis and 40% of patients with cirrhosis left education before achieving collage state exam, 9.1% of patients with pancreatitis and 25% of patients with cirrhosis had university education. Cirrhosis was diagnosed in 11 (16.7%) patients with chronic pancreatitis. In all patients, this was functional disorder Child-Pugh stage A, decompensation occurred in 54.5% of patients. Portal hypertension was diagnosed in 72.7% of patients and thrombosis of v. portae orv. lieanlis in 12 patients. Bleeding from gastroesophageal varices occurred in 1 patient with chronic pancreatitis, liver cirrhosis and v. portae thrombosis. Chronic pancreatitis was diagnosed in 2 (2.5%) patients with livercirrhosis. With respect to comorbidities, chronic obstructive bronchopulmonary disease and ischemic heart disease was significantly more frequent in patients with chronic pancreatitis. CONCLUSION: There are differences in social-behavioural background of both relatively common organ disorders. Chronic pancreatitis is more prevalent in people with lower education. The first contact with alcohol and regular abuse occur at younger age, smoking is also more frequent, with earlier onset. Both diseases rarely occur concurrently and simultaneous decompensation of both organs is not seen. We could speculate that early exposition to the known aetiopathogenic factors affect primarily pancreas. The rare concurrence of both diseases, particularly of their advanced stages, opposes all known theories.

Adipose tissue macrophages and atherogenesis - a synergy with cholesterolaemia.

Excessive LDL cholesterol concentration together with subclinical inflammation, in which macrophages play a central role, are linked pathologies. The process starts with the accumulation of macrophages in white adipose tissue and the switch of their polarization toward a pro-inflammatory phenotype. The proportion of pro-inflammatory macrophages in adipose tissue is related to the main risk predictors of cardiovascular disease. The cholesterol content of phospholipids of cell membranes seems to possess a crucial role in the regulation of membrane signal transduction and macrophage polarization. Also, different fatty acids of membrane phospholipids influence phenotypes of adipose tissue macrophages with saturated fatty acids stimulating pro-inflammatory whereas omega3 fatty acids anti-inflammatory changes. The inflammatory status of white adipose tissue, therefore, reflects not only adipose tissue volume but also adipose tissue macrophages feature. The beneficial dietary change leading to an atherogenic lipoprotein decrease may therefore synergically reduce adipose tissue driven inflammation.

May alcohol-induced increase of HDL be considered as atheroprotective?

It is well known that the consumption of moderate doses of alcohol leads to the increase of HDL-cholesterol (HDL-C). Atheroprotectivity of HDL particles is based primarily on their role in reverse cholesterol transport (RCT). In the study with a cross-over design 13 male volunteers were studied in two different regimens: i) drinking of 36 g alcohol daily and ii) drinking only non-alcoholic beverages, to test whether alcohol-induced increase of HDL cholesterol can affect cholesterol efflux (CHE) from cell culture of labeled human macrophages. Alcohol consumption induced significant (p < 0.05) increases of HDL cholesterol from 1.25 +/- 0.32 to 1.34 +/- 0.38 mmol/l and Apo A1 from 1.34 +/- 0.16 to 1.44 +/- 0.19 g/l. These changes were combined with a slight increase of cholesterol efflux from 13.8 +/- 2.15 to 14.9 +/- 1.85 % (p = 0.059). There were significant correlations between individual changes of HDL-C and Apo A1 concentrations and individual changes of CHE (0.51 and 0.60, respectively). In conclusion, moderate alcohol consumption changes the capacity of plasma to induce CHE only at a border line significance.

Life style change and reverse cholesterol transport in obese women.

HDL cholesterol resp. apolipoprotein A1 concentrations are tools to estimate individual CVD risk, although only a part of HDL particles participate in reverse cholesterol transport (RCT). This discrepancy was analyzed in life style change based on increase of physical activity and dietary counseling. Efflux of cholesterol from pre-labeled macrophages to plasma acceptors of tested individuals was used as an RCT measure. Changes of lipoprotein parameters, glucose, fasting insulin concentrations and RCT were analyzed in 15 obese women after 9-week intervention consisted of 5 sessions of increased physical activity per week. Controlled increase in physical activity for 9 weeks induced a decrease of body weight averaging 9 kg (ranged from 2.3 to 15.5 kg). The intervention leads to significant decreases of triglycerides, apoprotein A1 and apoprotein B concentration, whereas total cholesterol, LDL cholesterol and HDL cholesterol did not change significantly. The increase of RCT was not significant, but there was highly significant negative correlation between individual decrease of body weight and an increase of RCT. Significant increase of RCT was found in 13 persons with a weight reduction more than 3.5 kg. Substantial weight loss is necessary to increase RCT.

Actigenetic of ACE gene polymorphism in Czech obese sedentary females.

Obesity is a serious health problem worldwide and many genes have been implicated in determination of obesity, but our knowledge of the genes responsible for individual differences in weight loss after physical intervention are poor. One of the candidate genes is a gene for angiotensin-converting enzyme (ACE) ant its insertion/deletion (I/D) polymorphism. We have analyzed the association between the ACE gene variant in intervened obese females. Twenty four unrelated healthy obese (BMI > 29.9 kg/m(2), with abdominal type of obesity) premenopausal (age between 25 and 45 years) Czech Caucasian sedentary and non-diabetic females, pre-selected according the ACE I/D polymorphism (twelve II and twelve DD homozygotes) were studied in a medical research centre. They underwent 9 weeks intervention program (combination of the lowering of dietary intake to optimal level for the age and 3 times a week physical activity at fitness centre). The participants were supervised to sustain a heart rate of 65 % of maximum. Anthropometrical, biochemical parameters and body composition (Bodystat 1500) were analyzed before and after the intervention. Our study suggest, that in Czech Caucasian females I/D polymorphism within the ACE gene will have no major effect on weight loss. Interestingly, we have detected, that in obese females II genotype was associated with higher increase in basal metabolic rate (202 kcal per day) then in DD homozygotes (p<0.05), thus at least under some circumstances, this genetic variant may have an slight effect on BMI development.

Association between duration of the sleep and body weight.

Some studies have suggested that there could be an association between the duration of sleep in humans and development of the obesity. We have analyzed the group of the probands (n = 3970, 2038 males and 1932 females, aged 18-65 years), with permanent address in the Central or South Bohemia. We ascertained the relationship between the duration of their sleep (obtained per questionnaire) and body mass index, weight, height, the value of systolic and diastolic blood pressure, heart rate, waist and hip circumference, the values of total-, high density- and low density- cholesterol, thyroid hormone and body exercise performed. The optimal values of the body mass index (and optimal body weight) were associated with the duration of sleep 7 hours per night (P < 0.001). This association was found both in males and females and in both districts. Other anthropometrical and biochemical parameters were not associated with the sleep duration.

Molecular phylogenetic assessment of host range in five Dermanyssus species.

Given that 14 out of the 25 currently described species of Dermanyssus Dugès, 1834, are morphologically very close to each another, misidentifications may occur and are suspected in at least some records. One of these 14 species is the red fowl mite, D. gallinae (De Geer, 1778), a blood parasite of wild birds, but also a pest in the poultry industry. Using molecular phylogenetic tools we aimed to answer two questions concerning host specificity and synanthropicity: (1) is D. gallinae the only species infesting European layer farms?, and (2) can populations of D. gallinae move from wild to domestic birds and vice versa? Mitochondrial cytochrome oxidase I gene sequences were obtained from 73 Dermanyssus populations collected from nests of wild European birds and from poultry farms and these were analyzed using maximum parsimony and Bayesian inference. Mapping of the observed host range on the obtained topology and correlation with behavioural observations revealed that (1) host range is strongly dependent on some ecological parameters (e.g. nest hygiene, exposure to pesticides and predators), that (2) out of five species under test, synanthropic populations were found only in lineages of D. gallinae, and that (3) at least some haplotypes found in wild birds were very close to those found in association with domestic birds.

Double-blind, placebo-controlled evaluation of grass pollen specific immunotherapy with oral drops administered sublingually or supralingually.

Forty-one patients suffering from grass pollen allergy underwent specific immunotherapy with standardized allergen extract consisting of six grass pollens (H-Al per os) administered either sublingually or supralingually for one year. In order to investigate clinical and immunological changes induced by the administration of allergens via the oral mucosa, the double-blind, placebo-controlled, randomized design of the trial with 30 other patients enrolled in placebo groups was applied. Specific immunotherapy with oral drops administered sublingually or supralingually was performed in the same way, keeping the drops under or on the tongue, respectively, for 1-2 min before swallowing them; at the end of the trial the cumulative dose of the allergen was almost 20 times higher than that of the subcutaneous therapy with corresponding allergen preparation. Data about symptoms scores and drugs intake during grass pollen season, as well as skin reactivity, levels of specific IgG and IgE antibodies, before the study and after the study's completion, were obtained. It was found that both routes of administration are effective according to subjective clinical parameters and drug consumption, with a highly significant reduction of symptoms and drug intake favoring sublingual administration where a reduction of more than 60% was achieved. Only sublingual active group showed a significant increase in Dactylis glomerata-specific IgG serum levels. Adverse effects were limited to a small number of generally mild local and/or systemic reactions. The results suggest that the administration of allergens via the oral mucosa is safe and clinically effective, favoring the sublingual rather than supralingual route.

Pro-inflammatory gene expression in adipose tissue of patients with atherosclerosis.

Inflammatory changes, both in the arterial wall and adipose tissue, play a crucial role in the development of atherosclerosis. We measured the gene expression of tumor necrosis factor-alpha (TNFalpha), monocyte chemoattractant protein-1 (MCP-1), and interleukin 6 (IL-6) in adipose tissue (AT) of living kidney donors (LKD) and patients with peripheral arterial disease (PAD). Quantitative polymerase chain reaction (qPCR) and flow cytometry analyses were performed in subcutaneous (SAT), visceral (VAT), and perivascular adipose tissue (PVAT). Data of PAD patients showed significantly higher expression in VAT in all three genes (TNFalpha 5-fold, p<0.05; MCP-1 3.6-fold, p<0.05; IL-6 18.8-fold, p<0.001). The differences in PVAT and SAT were less significant. Total body pro-inflammatory status was documented by higher TNFalpha concentration in patients (4.86+/-1.4 pg/ml) compared to LKDs (2.14+/-0.9 pg/ml; p<0.001), as was hsCRP (11.8+/-7.0 in PAD; 1.5+/-0.48 in LKDs; p=0.017). We found no age-dependent relationship between gene expression vs. TNFalpha and hsCRP concentrations in both compared groups. No effect of the atherosclerosis score on gene expression and circulating inflammatory markers within the PAD group was observed. Our results suggest that the AT of PAD patients infiltrated with macrophages produces more cytokines involved in the development of inflammation and atherosclerosis.

Intake of carp meat from two aquaculture production systems aimed at secondary prevention of ischemic heart disease - a follow-up study.

Our previous study showed that a diet enriched with 400 g of carp per week improved plasma lipids in subjects after aortocoronary bypass (CABG). The aim of the present study is to determine whether the different carp farming systems have an impact on the effects of carp meat in secondary cardiovascular prevention. We examined 3 groups of patients after CABG over a 4-week period of spa treatment (108 persons, 73 males, 35 females, age over 60 years). We found no differences in baseline values of blood pressure or plasma lipids. The patients were given a standard spa diet (controls; N=36) or a diet enriched of 400 g of carp meat per week, enriched omega 3 (N=37) or cereal carp (N=35). Plasma lipid parameters were examined at start and after 4 weeks in a routine laboratory setting. Group consuming omega-3 carp showed the largest decline in total cholesterol, LDL cholesterol, triglycerides and an increase in HDL cholesterol (all p<0.01). We found that carp meat from the two production systems showed significantly different effects on plasma lipids. Further trials should be performed to clarify the exact causes of the differences.

Human adipose tissue accumulation is associated with pro-inflammatory changes in subcutaneous rather than visceral adipose tissue.

The importance of the involvement of adipose tissue macrophage subpopulations in obesity-related disorders is well known from different animal models, but human data are scarcer. Subcutaneous (n=44) and visceral (n=52) adipose tissues of healthy living kidney donors were obtained during living donor nephrectomy. Stromal vascular fractions were isolated and analysed by flow cytometry using CD14, CD16, CD36 and CD163 antibodies. Total macrophage numbers in subcutaneous adipose tissue increased (P=0.02) with body mass index (BMI), with a similar increase seen in the proportion of phagocytic CD14+CD16+CD36high macrophages (P<0.01). On the other hand, there was an inverse correlation between anti-inflammatory CD14+CD16-CD163+ macrophages (P<0.05) and BMI. These correlations disappeared after excluding obese subjects (BMI ⩾30 kg m-2) from the analysis. Interestingly, none of these subpopulations were significantly related to BMI in visceral adipose tissue. Obesity per se is associated with distinct, highly phagocytic macrophage accumulation in human subcutaneous adipose tissue.

Adipose tissue and atherosclerosis.

High-energy intake which exceeds energy expenditure leads to the accumulation of triglycerides in adipose tissue, predominantly in large-size adipocytes. This metabolic shift, which drives the liver to produce atherogenic dyslipidemia, is well documented. In addition, an increasing amount of monocytes/macrophages, predominantly the proinflammatory M1-type, cumulates in ectopic adipose tissue. The mechanism of this process, the turnover of macrophages in adipose tissue and their direct atherogenic effects all remain to be analyzed.

The effect of ectopic fat on graft function after living kidney transplantation.

Renal transplantation is associated with a large number of risk factors that can have an influence on early renal graft function (ERGF). One of these factors could be the increasing number of obese kidney donors. The mechanisms of reduced ERGF in obese kidney donors are still poorly understood. To that end, we compared ERGF in recipients with body mass index (BMI), perivascular fat and plasma inflammation markers of live kidney donors. We hypothesized that the BMI of donors would negatively correlate with an average increase of glomerular filtration rate (GFR) and that it would also be associated with increased perivascular and plasma inflammation markers in the first seven days after transplantation. Between January 2013 and December 2014, some 58 living kidney transplantation pairs were included in the study. Donor and recipient demographic data, preoperative BMI, blood C-reactive protein (CRP) and adiponectin levels, perivascular adipose tissue (PAT) samples and recipient blood creatinine levels were analyzed. The median CRP of donors was 0.68 mg/l (max: 8.66 mg/l, min: 0.33 mg/l), the median of M1 macrophages (CD14+CD16+) in one gram of PAT was 5940 (max: 41 100, min: 248) and the median of adiponectin was 411 930 pg/ml (max: 14 217 000, min: 167 300) in plasma. We did not find any association between early renal graft function and the percentage of M1 macrophages in donor perirenal adipose tissue (p=0.83, r=0.03, n=58), adiponectin (p=0.65, r=0.06, n=58) or CRP (p=0.16, r=0.2, n=58) in plasma. The obesity level of donors, expressed as BMI, did not correlate with early renal graft function in the first seven days after transplantation. The associations between ERGF and plasma and perivascular fat inflammation markers were not significant. We confirmed a negative correlation between the BMI of recipients and an average increase of GFR in the first seven days after transplantation (p<0.02, r=-0.325, N=58). We confirmed a negative correlation of adiponectin plasma concentration to the BMI of donors.

Macrophage phenotypes in the adipose tissue of postmenopausal women.

Atherosclerosis pathology is the interplay between high intravascular LDL particle concentration and monocyte/macrophage presence within the sub-endothelial space of the artery. In this project, phenotypes of macrophages connected with subclinical inflammation in adipose tissue of living kidney donors were studied. Samples of subcutaneous adipose tissue of living kidney donors (n=36) were exposed to collagenase. Stromal vascular fraction (SVF) was eluted from the samples, then labeled with monoclonal antibodies (anti-CD14 and anti-calprotectin), conjugated with fluorochromes and analyzed by flow cytometry. The positive correlation between the number of total macrophages and calprotectin-positive macrophages with BMI in the subcutaneous adipose tissue of postmenopausal women was demonstrated (p<0.05; R=0.43 and p<0.01; R=0.60), whereas no positive correlation in premenopausal women and men was shown. In conclusion, we documented a significant effect of BMI increase on the presence of total macrophages in adipose tissue of postmenopausal women, in contrast to premenopausal women. This difference was much more pronounced when proinflammatory macrophages with membrane-bound calprotectin were analyzed.

Is the amount of coronary perivascular fat related to atherosclerosis?

Interesting and stimulating data about the effect of the perivascular adipose tissue size on atherogenesis are based mainly on CT findings. We studied this topic by directly analyzing perivascular adipose tissue in explanted hearts from patients undergoing transplantation. Ninety-six consecutive patients were included, including 58 with atherosclerotic coronary heart disease (CHD) and 38 with dilation cardiomyopathy (DCMP). The area of perivascular fat, area of the coronary artery wall, and ratio of CD68-positive macrophages within the perivascular fat and within the vascular wall were quantified by immunohistochemistry. There was no significant difference in the perivascular adipose tissue size between the two groups. Nevertheless, there was a significantly higher number of macrophages in the coronary arterial wall of CHD patients. In addition, we found a close relationship between the ratio of macrophages in the arterial wall and adjacent perivascular adipose tissue in the CHD group, but not in the DCMP group. According to our data interaction between macrophages in the arterial wall and macrophages in surrounding adipose tissue could be more important mechanism of atherogenesis than the size of this tissue itself.