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In contrast to the decreasing burden related to cardiovascular disease (CVD), the burden related to dysglycemia and adiposity complications is increasing in Czechia, and local drivers must be identified. A comprehensive literature review was performed to evaluate biological, behavioral, and environmental drivers of dysglycemia and abnormal adiposity in Czechia. Additionally, the structure of the Czech healthcare system was described. The prevalence of obesity in men and diabetes in both sexes has been increasing over the past 30 years. Possible reasons include the Eastern European eating pattern, high prevalence of physical inactivity and health illiteracy, education, and income-related health inequalities. Despite the advanced healthcare system based on the compulsory insurance model with free-for-service healthcare and a wide range of health-promoting initiatives, more effective strategies to tackle the adiposity/dysglycemia are needed. In conclusion, the disease burden related to dysglycemia and adiposity in Czechia remains high but is not translated into greater CVD. This discordant relationship likely depends more on other factors, such as improvements in dyslipidemia and hypertension control. A reconceptualization of abnormal adiposity and dysglycemia into a more actionable cardiometabolic-based chronic disease model is needed to improve the approach to these conditions. This review can serve as a platform to investigate causal mechanisms and secure effective management of cardiometabolic-based chronic disease.

Introduction: Despite the continuously growing number of therapeutic options for type 2 diabetes mellitus (T2DM) including insulins, a large percentage of patients fail to achieve HbA1c targets. Several real-world studies focused on patients with T2DM receiving insulin treatment in outpatient settings were conducted, but information about real-world in-hospital insulin management is lacking. The aim of this study was to describe the management of insulin therapy with a focus on basal-bolus and premixed insulin regimens in patients with T2DM under routine in-hospital medical practice in the Czech Republic., Methods: This non-interventional prospective study was conducted from June 2014 to December 2017 in 22 centers in the Czech Republic under routine clinical practice conditions. Adult patients admitted to hospital with metabolically uncontrolled T2DM [HbA1c ≥ 60 mmol/mol; > 7.6% Diabetes Control and Complications Trial (DCCT)] and there treated with basal-bolus and premixed insulin regimens were documented during hospitalization., Results: Overall, 369 patients with T2DM (54.7% male, mean age 64.44 ± 13.84 years, BMI 31.10 ± 6.00 kg/m 2 , duration of diabetes 8.11 ± 9.93 years, HbA1c 95.90 ± 24.38 mmol/mol, length of stay was 7.94 ± 4.53 days) were included. The percentage of glucose values under 10 mmol/l at time of randomization (the group with basal-bolus insulin regimen vs. the premix insulin regimen group) was 24.2% vs. 33.5% (p = 0.053), at time of first insulin dose adjustment it was 43.1% vs. 50.0% (p = 0.330), and 1 day before hospital discharge it was 61.7% vs. 61.4% (p = 0.107). A hypoglycemic event occurred in a total of 15 patients in the basal-bolus regimen group, and no hypoglycemic event occurred in the premixed insulin regimen group., Conclusion: In-hospital insulin management regarding basal-bolus and premixed insulin regimens is safe and in concordance with current international recommendations.

Arterial hypertension is one of the most frequent comorbidities of patients with type 2 diabetes, the rates are approximately three times higher among diabetics than in general population. The presence of both diseases increases the cardiovascular risk of the patients two to three times. Therefore, it is essential to control the blood pressure values vigorously down to 130/80 mmHg as recommended in guidelines of the Czech Hypertension Society. According to EHES Study data, the blood pressure control is not sufficient in Czech diabetic population. The target values are reached in only 29 % of diabetics. A well organized system of care, intensive antihypertensive treatment, self-monitoring of patients or appointment reminder system can be used to improve blood pressure control.

Cushing´s syndrome is a rare disease with the population prevalence about 40 patients per 1 million inhabitants, the number among patients with diabetes mellitus (DM) is probably higher. The screening of CS among the DM patients showed the prevalence between 0-2.9% in the groups of outpatients and 2-2.9% among the patients admitted to the hospital at the time of randomization. The routine screening of CS among patients with type 2 DM is not recommended at the moment. Nevertheless, the possibility of CS should be taken into the account mainly in patients with hypertension, obesity, microvascular complications, bad level of metabolic control or high insulin doses, especially, if more of these features are present. A hormonal profile of patients with adrenal incidentaloma should be evaluated.

Aims/hypothesis: The need for understanding obesity as a chronic disease, its stigmatization, and the lack of actionability related to it demands a new approach. The adiposity-based chronic disease (ABCD) model is based on adiposity amount, distribution, and function, with a three stage complication-centric rather than a body mass index (BMI)-centric approach. The prevalence rates and associated risk factors are presented., Methods: In total, 2159 participants were randomly selected from Czechia. ABCD was established as BMI ≥ 25 kg/m 2 or high body fat percent, or abdominal obesity and then categorized by their adiposity-based complications: Stage 0: none; Stage 1: mild/moderate; Stage 2: severe., Results: ABCD prevalence was 62.8%. Stage 0 was 2.3%; Stage 1 was 31.4%; Stage 2 was 29.1%. Comparing with other classifiers, participants in Stage 2 were more likely to have diabetes, hypertension, and metabolic syndrome than those with overweight, obesity, abdominal obesity, and increased fat mass. ABCD showed the highest sensitivity and specificity to detect participants with peripheral artery disease, increased intima media, and vascular disease., Conclusion/interpretation: The ABCD model provides a more sensitive approach that facilitates the early detection and stratification of participants at risk compared to traditional classifiers., Competing Interests: Dr. Mechanick has received honoraria for lectures and program development from Abbott Nutrition International. The other authors have not conflict of interest., (© 2021 The Authors. Obesity Science & Practice published by World Obesity and The Obesity Society and John Wiley & Sons Ltd.)

Glucagon is actually used in the treatment of severe hypoglycemia. Due to technological difficulties with stability in solution, glucagon in currently available preparations was contained in a form of powder, which needs to be first dissolved in the water before application. This particular administration of glucagon was associated with a certain delay and several mistakes that decreased real amount of glucagon injected in the body. Currently two new formulas of glucagon appeared on the market - first as a stable liquid solution, second as a powder for nasal administration. This article summarizes currently available information about these preparations.

Latent Autoimmune Diabetes in Adults (LADA) is an autoimmune disease arising at adulthood. LADA is characterized by a less intensive autoimmune process, slower progression and a mild metabolic decompensation at onset compared with young-onset type 1 diabetes mellitus. The onset of LADA is usually in non-obese patients over 30, without prominent features of metabolic syndrome and insulin resistance. Nevertheless it may be falsely classified as type 2 diabetes, especially, when diagnosed in older age and for the possibility of non-insulin treatment for at least 6 months after diagnosis. LADA is treated early with insulin and combined with metformin in patients with a higher level of insulin resistance. Clinical studies suggested also effectivity of other oral antidiabetics enabling preservation of residual β-cell function, such as particularly incretines.

Alcohol is an important risk factor for development of numerous chronic diseases and it is a cause of 3 million deaths worldwide every year. The Czech Republic is one of the countries with highest alcohol consumption per capita in the world. AUDIT and CAGE are examples of fast and easy screening tools for early diagnostics of patients at risk that help in estimating a level of risk related to patients drinking. The screening is followed by simple advice for risk drinking and brief intervention for harmful drinking, based on the estimated level of risk. This approach is an easy and effective prevention measure to reduce risk and harmful alcohol drinking and prevent further progression of drinking. Implementation of this approach into routine clinical practice can be a significant step in reducing alcohol consumption in the Czech population.

Aims: The aim of this study is to establish the prevalence of diabetes (DM) and prediabetes in the Czech population aged 25-64 years and to evaluate the relationships with various cardiometabolic, sociodemographic, and lifestyle risk factors., Methods: This was an epidemiological study with a stratified, crosssectional, random sampling design. Sociodemographic, lifestyle, and anamnestic data were collected through interviewer-administered questionnaires, medical examination, and biochemical assays., Results: Among the 1189 participants, 114 were diagnosed with DM (9.6%), 330 with prediabetes (27.8%) and 745 were non-diabetes/non-prediabetes individuals (62.7%). Logistic regression analysis showed that overweight, general and abdominal obesity, hypertension, and lower level of HDL (increased risk) significantly increased the risk of both prediabetes and DM, while living in the cities diminished risk of DM. Among lifestyle variables the significant increased risk of prediabetes and DM was found for smokers and ex-smokers. In other lifestyle variables (marijuana lifetime prevalence, physical activity and frequency of alcohol drinking) the significantly higher or lower risk for prediabetes or DM was not found., Conclusions: The study shows a high prevalence of DM and prediabetes in the Czech population of age between 25 and 64, providing data on their association with several risk factors., Competing Interests: Declaration of Competing Interest The authors declare that they have no conflict of interest., (Copyright © 2020 Elsevier B.V. All rights reserved.)

Objective: The aim of this article is to present a summary of the actual diagnostic possibilities and differentiation of MODY (Maturity-Onset Diabetes of the Young) from gestational diabetes (GDM) found during routine screening, and specific aspects of care and treatment of MODY during pregnancy and early postpartum period., Design: Rewiev., Settings: Centre for Research of Diabetes, Metabolism and Nutrition; Second Department of Internal Medicine University Hospital Královské Vinohrady and Third Faculty of Medicine, Prague. Department of Internal Medicine, Second Faculty of Medicine, Charles University, Prague., Methods: Recent publications selected in PubMed with the key words MODY, gestational diabetes., Results: Many patients with MODY, especially the glucokinase MODY, can be first diagnosed during pregnancy. It is estimated that MODY patients account for up to 5% of GDM cases found in routine screening of GDM. MODY should be considered in lean women around 25 years of age, with a positive family history of diabetes in one of the parents. The differentiation of MODY from GDM is of particular importance not only for the different management and goals of antidiabetic therapy and planning ultrasound controls of fetal growth during pregnancy, but also because of the risk of hyperinsulinemic hypoglycemia in newborns., Conclusion: Recognition of MODY during pregnancy and adherence to existing recommendations concerning specific care of these patients is essential for the optimal course of their pregnancy and proper care of the newborn in the early postpartum period.

Purpose: Screening of gestational diabetes/GDM (although different in different countries) represents a standard procedure allowing to identify women with pregnancy-associated diabetes. Some of the women with GDM (up to 5%) may, however, suffer from previously undiagnosed MODY (Maturity-Onset Diabetes of the Young). Currently, no international or local guidelines focused on the identification of MODY among GDM exist. Thus, the aim of this manuscript is to propose a clear guide for clinicians on how to detect MODY among pregnant women with gestational diabetes., Methods: Based on the available literature about diagnosis (in general population) of MODY and management of MODY (both, in general population and in pregnant women), we propose a clear clinical guide on how to diagnose and manage MODY in pregnancy., Results: The manuscript suggests a feasible clinical approach how to recognize MODY among patients with GDM and how to manage pregnancy of women with three most common MODY subtypes., Conclusion: A correct classification of diabetes is, nonetheless, essential, particularly in case of MODY, as the management of pregnant women with MODY is different and the correct diagnosis of MODY enables individualized treatment with regard to optimal pregnancy outcomes.

Mobile and wearable technologies offer patients with diabetes mellitus new possibilities for data collection and their more effective analysis. The Diabesdagboga smartphone application and the Diani web portal enable to collect and analyze glycaemia values, carbohydrates intake, insulin doses and the level of physical activity. The data are not only accessible in the corresponding smartphone but also automatically transferred to an Internet portal, where they may be completed by the records from an electronic pedometer and continuous glucose monitor. All these data may then be displayed in various types of graphical outputs and are available to both the patient and the physician. The case report of a patient who has used the system for almost two years shows a significant improvement in metabolic compensation (a decrease in the mean HbA1c value by 18.6 mmol/mol as compared with the previous period).

Lifetime prevalence of illicit drug use is high in the Czech Republic (CR) - it reached 31.2 % of the population in the age group 15-64 years in 2017. Diabetes mellitus (DM) applies to 8.8 % of the Czech population. Risks of illicit drug use in diabetic patients are related both to their effect on glucose metabolism and to the lifestyle associated with illicit drug use, which might lead to worsened glycemic control and increased morbidity and mortality of the patients. Cannabis use, being the most common illicit drug use in the CR, presumably does not have a direct effect on glucose metabolism. However, it increases appetite and decreases self-control. Opioids/opiates disrupt insulin secretion, which consequently leads to hyperglycaemia. On the other hand, hypoglycaemia might be an adverse effect of opioid treatment in diabetic patients. Cocaine and other stimulants such as MDMA increase a blood glucose level and increase the risk of diabetic ketoacidosis in Type 1 DM. Patients with DM who use illicit drugs should therefore be sufficiently informed about health risks related to illicit drug use in combination with DM.

The results of two large multicentric, observational and cross-sectional studies (the DIAINFORM study and BALI study) reflecting routine clinical practice for insulin treatment in type 2 diabetes mellitus in the Czech Republic were published recently. In this commentary, we analyze these results and compare them with the findings of similar studies conducted in other countries within the last decade. The analysis focuses on achievement of glycated hemoglobin goals, insulin dosage and frequency of hypoglycemia.

Alcoholic drinks are one of the risk factors for hypoglycemia. Ethanol inhibits gluconeogenesis, decreases a level of growth hormone and impairs hypoglycemia awareness. The risk of hypoglycemia while drinking alcohol can be reduced by parallel ingestion of food (saccharides). Some recommendations also mention the change of insulin doses.

Hypoglycemia particularly severe one is important side effect of diabetes therapy with impact on patient´s quality of life and mortality. The highest risk of hypoglycemia is connected with insulin therapy followed by derivates of sulfonylurea (SU) and glinides. The risk of hypoglycemia found in observational studies were 2-3 times higher in patients treated with SU and 3-4 higher when treated with insulin compared with other types of antidiabetics. The risk of hypoglycemia is increased in patients over 75 years of age, with longer period of treatment with insulin and in those treated with several types of antidiabetics.

Despite the continuously improving treatment options, many patients with type 1 (T1DM) and type 2 diabetes (T2DM) still do not achieve the recommended treatment goals. The article provides summary and commentary of the results of DIAINFORM study focused on the level of metabolic control in T1DM and T2DM patients treated with insulin in the Czech and Slovak Republics. The overall percentage of patients with HbA1c 3 mmol/mol in the T1DM group was 29.9 % and in the T2DM group was 33.4 %.

Hypoglycemia is a side effect of the therapy primarily with insulin, sulphonylurea derivates and glinides. Its therapy is based on the immediate ingestion of sacharides, preferably glucose. Amount of 15-20 g is recommended as its optimal dose, although several recent studies are suggesting amount related to the patient´s weight. The therapy of severe hypoglycemia in the non-professional settings is based on glucagon injection, in the professional ones intravenous administration of glucose is preferable option.

Recurrent, unexplained, eventually severe hypoglycemias in patients with type 1 diabetes mellitus (T1DM) may be rarely but yet associated with the onset of adrenal insufficiency (AI), present in approximately 0.5 % of patients with T1DM, appearing usually several years after diabetes diagnosis, usually in middle age and more frequently in women. Screening for AI in patients who complained of recurrent hypoglycemias, difficult to explain by common causes, was ineffective and therefore it is not recommended. Nevertheless, the possibility of manifestation of AI in connection with above mentioned cases in patients with T1DM should be taken into consideration and usual symptoms of AI such as weakness, lack of appetite, weight loss, orthostatic hypotension, hyperpigmentation or ions disorders should be examined.

Introduction: The aim of the study was to determine the level of metabolic control in type 1 diabetes mellitus (T1DM) and type 2 diabetes mellitus (T2DM) patients in the Czech and Slovak Republics., Methods: A non-interventional prospective (observational) study was conducted from January 2015 until April 2016 in routine clinical practice settings at 141 centers in the Czech and Slovak Republics. Data were analyzed from a total of 425 patients with T1DM and 1034 patients with T2DM, proportionally corresponding to the number of patients in both countries. The primary objective of the study was to determine the percentage of patients with HbA1c < 7% (53 mmol/mol)., Results: Patients with T1DM: In this group of patients (55.8% males, mean age 45.9 ± 14.83 years, BMI 25.8 ± 4.21 kg/m², diabetes duration 12.1 ± 9.44 years), 29.9% reached HbA1c levels < 53 mmol/mol. Patients with T2DM: In this group of patients (50.3% male, mean age 63.9 ± 9.65 years, BMI 31.0 ± 5.19 kg/m², diabetes duration 12.4 ± 7.47 years, duration of insulin therapy 5.8 ± 4.71 years), 33.4% reached HbA1c levels < 53 mmol/mol., Conclusion: The overall percentage of patients with HbA1c < 53 mmol/mol in the T1DM group was 29.9% and in the T2DM group was 33.4%. Despite an increasing number of treatment options, most patients still fail to reach the recommended HbA1c targets., Funding: Sanofi, Czech Republic.

The discovery of MODY (Maturity-Onset Diabetes of the Young) and the elucidation of its heritability enabled more precise clinical characteristics of different MODY subtypes and led to understanding that glucokinase MODY (GCK-MODY) is not associated with vascular complications in long term follow-up, whereas MODY of transcription factors (e.g. HNF1A-MODY) is in case of bad metabolic control connected with the acceleration of particularly microvascular complications. There is a strong evidence of the needlessness of any specific antidiabetic treatment in prognosticaly favourable GCK-MODY (except for the pregnancy). On the contrary, in MODY of transcription factors, including the most common one - HNF1A-MODY, the treatment, traditionally based on sulphonylurea derivatives (and after their failure on insulin) is required. Due to wider spectrum of available antidiabetic agents offering individualization of the treatment, the question of efficacy of other antidiabetic agents in MODY patients arises. This review article summarizes current knowledge of therapeutic options in patients with MODY.Key words: insulin - MODY - oral hypoglycemic drugs - treatment.

Maturity Onset Diabetes of the Young (MODY) comprises inherited forms of diabetes mellitus caused by the mutations in the genes involved in the development, differentiation and function of beta-cells. The majority of patients with MODY remains misdiagnosed and erroneously classified as type 1 or type 2 diabetic patients. Correct MODY diagnosis is, however, essential since it enables individualization of treatment, assessment of the prognosis and identification of diabetes among patient´s relatives. Clinical presentation of MODY is highly variable and it could resemble other types of diabetes, thus identification of MODY patients might be difficult. In this review, we describe typical clinical presentation of the most common MODY subtypes, we summarize current diagnostic guidelines in confirmation of MODY and we raise the question of possible need for extension of current clinical criteria indicating a patient for molecular-genetic testing.Key words: clinical course - diagnosis - differential diagnosis - glucokinase - hepatocyte nuclear factors - MODY.

Introduction: Moderate to medium decrease in glomerular filtration (GFR) in individuals with chronic kidney disease (CKD) does not need to be associated with hyperphosphatemia due to an adaptive decrease in tubular reabsorption of phosphates (TRPi) in residual nephrons. The clinical assessment of this function is performed based on the measurement of fractional phosphate excretion (FEPi), which is a quantity specifying the proportion of the filtered amount of phosphates which is excreted in the urine. This quantity may provide useful information about the involvement of kidneys in phosphate homeostasis of the internal environment. This study focuses on the comparison of a kr(FEPi) value examined based on a ratio of a phosphate clearance (CPi) and a creatinine clearance (CKr) marked kr(FEPi), and a value calculated based on a ratio of CPi and an exactly measured GFR as an inulin clearance (Cin), marked as in(FEPi).The goal of comparing the two methods of examining FEPi was to establish to what extent it is possible to evaluate the degree of inhibition of tubular phosphate transport in residual nephrons based on a simple examination of kr(FEPi) ., Methodology: The examination of in(FEPi) and kr(FEPi) was carried out for 53 patients with CKD. The values of the examined quantities were as follows: SKr 199±45 µmol/l; SPi 1.41±0.29 mmol/l; CKr 0,95±0.36 ml/s/1.73 m2; Cin 0.71±0.25 ml/s/1.73 m2. For the purpose of comparison a cohort of 18 healthy volunteers was examined., Results: For individuals with CKD an average value of kr(FEPi) equalled 29.1±10.9% and in(FEPi) 52.4±4.3%. The values of in(FEPi) were higher than kr(FEPi) (p<0.001) for all patients, although an average CPi value for patients with CKD did not significantly differ from the control cohort (0.22 vs 0.21 ml/s/1.73 m2). The values of in(FEPi) increased proportionally to SKr values and at higher values SKr (>300 µmol/l) they gradually approached 100% (indicating the complete inhibition of tubular reabsorption of phosphates in residual nephrons). The values of in(FEPi) were higher in all patients with CKD than kr(FEPi) as expected, likely because the value CKr decreases at a slower rate than Cin (GFR) in individuals with CKD as a result of increased tubular secretion of creatinine in residual nephrons., Conclusion: The results of this study support the assumption that, provided the values of kr(FEPi) which are easily measurable in clinical practice have reached 50-60%, almost complete inhibition of tubular reabsorption of phosphates in residual nephrons must be assumed and no favourable effect of phosphatonins on renal phosphate excretion can be expected. When looking for new possibilities of inhibition of tubular phosphate reabsorption, potential adverse effects of phosphatonins on organs must be considered.

Tubular transport of sodium (TNa+) and chloride (TCl-) is decreased in patients with chronic kidney disease. The decrease of TCl- is relatively lower than that of TNa+. These changes of tubular transport of Na+ and Cl- participate on the development of acid base disturbance in patients with chronic kidney disease and with their glomerular filtration rate lower than 0.5 ml/s/1.73 m2.

Background: The protein tyrosine phosphatase nonreceptor type 22 (PTPN22) gene encodes lymphoid tyrosine phosphatase (LYP), which is expressed primarily in lymphoid tissues. The functional but geographically highly variable PTPN22 single-nucleotide polymorphisms (SNPs), particularly c.1858C>T, contribute to the onset and progression of autoimmunity-associated diseases and facilitate the expression of disease-associated autoantibodies. In Central Europe, 17-25% of patients with monogenic diabetes (maturity-onset diabetes of the young, MODY) transiently express islet cell autoantibodies., Methods: We addressed the links between the functional and geographically variable PTPN22 SNPs with MODY manifestation and the expression of islet cell autoantibodies in 276 MODY patients who originated from four regions (the Czech Republic, Israel, Japan and Brazil)., Results: The frequency of PTPN22 polymorphisms in the MODY patients was similar to those in geographically matched healthy populations, with the exception of c.788G>A, the minor allele frequency of which was significantly elevated in the Czech hepatocyte nuclear factor 1-α (HNF1A) MODY patients [odds ratio (OR) 4.8, 95% confidence interval (CI) 2.2-10.7] and the Brazilian MODY patients (OR 8.4, 95% CI 1.8-39.1). A barely significant increase in the c.788G>A minor allele was also detected in the islet cell autoantibody-positive Czech MODY patients. However, c.788A behaves as a loss-of-function mutant in T cells, and thus protects against autoimmunity., Conclusions: MODY patients (including islet cell autoantibody-positive cases) do not display any increase in autoimmunity-associated PTPN22 alleles. The absence of autoimmunity-associated PTPN22 alleles was also demonstrated in latent autoimmune diabetes in adults, which suggests that the slow kinetics of the onset of autoantibodies is subject to a regulation that is different from that experienced in type 1 diabetes and other autoimmune disorders., (© 2015 S. Karger AG, Basel.)

Insulin secretion in patients with manifested diabetes mellitus tends to disappear months to decades after the diagnosis, which is a clear sign of a gradual loss of pancreatic islet beta-cells. In our sample of 30 type 2 diabetic patients, whose disease manifested between 30 and 45 years of age, about a half have retained or even increased insulin secretion 30 years later, while the other half exhibit a much diminished or lost insulin secretion. Factors that can damage or destroy beta-cells can be divided into the following groups: Metabolic factors: hyperglycemia and glucotoxicity, lipotoxicity, hypoxia, reactive oxygen species; Pharmacological factors: antimicrobial medication pentamidine, SSRI antidepressants; Factors related to impaired insulin secretion: MODY type diabetes; Environmental toxic factors: rat poison Vacor, streptozotocin, polychlorinated and polybrominated hydrocarbons; Disorders of the exocrine pancreas: tumor infiltration, fibrous infiltration, chronic pancreatitis, cystic fibrosis; Infections, inflammation, autoimmunity, viral factors: Coxsackie viruses, H1N1 influenza, enteroviruses. We are currently working on finding other factors leading to beta-cell damage, studying their effect on apoptosis and necrosis and looking for possible protective factors to prevent this damage. We our increasing knowledge about the mechanisms of beta-cell damage and destruction we come ever closer to suggest measures for their prevention. In this review we offer a brief and simplified summary of some of the findings related to this area.Key words: pancreatic islet beta-cells of Langerhans - factors damaging or destroying beta-cells - insulin secretion.

Background: The aim of the study was to evaluate the in vitro quality of cryopreserved red blood cells obtained from different sources with or without leucodepletion and stored at 4±2 °C in AS-3 for up to 21 days., Materials and Methods: Red blood cells were collected by four methods: double erythrocytapheresis, whole blood collection with buffy coat removal, double erythrocytapheresis with in-line leucofiltration, or whole blood collection with in-line leucofiltration. All four types of red blood cells were frozen in 40% glycerol after collection and stored at a temperature below -65 °C for at least 30 days, thawed, deglycerolised and subsequently reconstituted in AS-3. The in vitro haematological and biochemical properties of the thawed red blood cells were tested on days 0, 7, 14, and 21 after deglycerolisation and reconstitution., Results: Overall, 72 units were processed. Leucodepletion of cryopreserved red blood cells units reduced haemolysis, lowered ammonia concentration, preserved pH and osmolality and led to sustained higher concentrations of ATP. In contrast, the source of red blood cells (apheresis or whole blood) did not affect their quality., Discussion: The quality of all investigated red blood cells units was the same as or even better than that of erythrocytes obtained from double erythrocytapheresis with a 24-hour survival of at least 86% after up to 3 weeks of storage in AS-3.

It was recently suggested that routine islet cell autoantibody testing should be performed to discriminate maturity-onset diabetes of the young (MODY) from type 1 diabetes mellitus (T1DM). This is the first report ever to describe the familial manifestation of T1DM autoimmunity in nonobese HNF1A-MODY subjects and the presence of islet antigen-2 (IA-2) antibodies in MODY subjects. Three nonobese subjects in an age range of 14-35 years were diagnosed with HNF1A-MODY (p. Arg159Gln mutation). All the tested subjects had detectable (but varying) levels of islet cell autoantibodies (i.e., antibodies against glutamate decarboxylase or IA-2) in the absence of other T1DM characteristics. They displayed long-term expression of intermediate fasting C-peptide levels, ketoacidosis was absent even in periods of spontaneous insulin withdrawal, and full dependence on externally administered insulin was not detected in any of them although better glycemic control was achieved when insulin was supplemented. The course of the disease was similar to that of the autoantibody-negative HNF1A-MODY subjects. The case questions the selectivity of autoantibodies as a marker of T1DM or late-onset autoimmune diabetes of adulthood (LADA) over MODY and challenges the use of autoantibodies as a universal negative marker of MODY in an effort to decrease the cost of health care, as it may eventually lead to the wrong diagnosis and thus to the incorrect treatment. Further research should involve examination of the autoantibody titers and prevalence in large and geographically diverse cohorts of MODY subjects selected for genetic testing (regardless of their autoantibody titers) as well as determination of the islet cell autoantibody kinetics in the course of MODY onset and progression., (Copyright © 2013 S. Karger AG, Basel.)

Introduction: Macleaya microcarpa (Papaveraceae family) has been of considerable interest in recent years as a prospective source of quaternary benzo[c]phenanthridine alkaloids (QBAs) related to many pharmaceutical beneficial effects. For this purpose, a quantitative, efficient and fast method to isolate the QBAs from the plant material is required., Objective: To optimise and compare pressurised liquid extraction (PLE) with Soxhlet extraction and maceration in order to estimate extraction conditions for fast and efficient isolation of QBAs contained in the roots of Macleaya microcarpa., Methodology: The QBAs were extracted by PLE, Soxhlet extraction and maceration at different conditions (solvent, time, etc.). Reversed phase HPLC with diode-array detector was utilised for their determination and quantification. To optimise the PLE procedure, the variable parameters, including temperature (40-150 °C), sample-to-inert material ratio, extraction time (5-30 min) and number of extraction cycles (1-4), were also tested., Results: Quantitative determination of QBAs resulted in 0.2-2.8 mg/g, 0.3-2.5 mg/g and 0.3-3.1 mg/g for PLE, Soxhlet extraction and maceration. To produce the yields mentioned above, PLE required only up to 30 min compared with 21 h for Soxhlet extraction and 49 days for maceration., Conclusion: PLE provided an effective and fast extraction of QBAs from M. microcarpa roots and can be recommended as an alternative isolation method to conventional techniques for QBAs from the plant sources., (Copyright © 2012 John Wiley & Sons, Ltd.)

Macleaya microcarpa (Maxim.) Fedde belongs to the genus Macleaya, family Papaveraceae. Together with the better known and more frequently studied species M. cordata (Willd.) R. Br. it is a main source of quaternary benzo[c]phenanthridine alkaloids. Using HPLC we determined the content of eight isoquinoline alkaloids in the aerial and underground parts of 1-, 2-, 12- and 13-year old plants and followed their changes during the vegetative period. The dominant alkaloid of all samples collected in the end of this period was allocryptopine (3.8-13.6 mg/g for aerial parts, 24.2-48.9 mg/g for underground parts). Chelerythrine, sanguinarine and protopine were also present in both parts of the plant. Additionally, measurable concentrations of chelilutine (CL), chelirubine (CR), macarpine (MA) and sanguirubine (SR) were detected in underground parts. The most important finding was that contents of CR, CL, SR and MA in the 12- and 13-year old plant roots were significantly higher (approximately 3-fold for CR, 6-fold for CL, 5-fold for SR, and at least 14-fold for MA) than in 1- or 2-year old plants. The proportion of individual alkaloids in aerial and underground parts thus changed significantly during the vegetative period.

Background: The Czech Republic decided to build the frozen red blood cell (RBC) bank for military contingency and civil emergency preparedness. Storage methods were validated with (51)Cr recovery measures out to 21 days postthaw., Study Design and Methods: A total of 139 apheresis RBC units collected in additive solution (additive solution 3 [AS-3] or saline-adenine-glucose-mannitol [SAG-M]) were obtained from volunteers by double erythrocytapheresis; all were sterilely glycerolized, frozen, stored for at least 30 days at a minimum of -65 degrees C in collection or cryogenic bags, thawed, sterilely deglycerolized, and stored at 4 +/- 2 degrees C for up to 21 days in AS-3 or SAG-M. Units were sampled before storage and after thaw weekly. Metabolic intermediates and (51)Cr recovery measures were obtained to determine the index of therapeutic effectiveness (ITE)., Results: Despite losses associated with cryoconservation and eventual transfer to cryogenic bags, 3-week storage with optimum ITE was obtained with frozen storage in primary collection or cryogenic bags and postthaw storage in AS-3. Such cells would pass as acceptable in vitro hematology and biochemistry variables., Conclusions: A system for frozen RBC storage with 3-week, postthaw, liquid storage has been validated. With commercially available components and automated processing, it is capable of supporting civilian rare blood and emergency and international military combat and peacekeeping support use patterns.

The spectral, especially fluorescence properties, of seven selected quaternary benzo[c]phenantridine alkaloids (sanguinarine, chelerythrine, chelirubine, sanguirubine, chelilutine, sanguilutine, and macarpine) were studied in presence and in absence of double-stranded DNA. This study has proved dramatic differences in fluorescence emission of all studied alkaloids in presence of calf thymus DNA in comparison to fluorescence of free alkaloids. The most remarkable are changes in emission spectra of macarpine, chelirubine, and sanguirubine. Association constants (logK) for interaction of all studied alkaloids with CT DNA were calculated.

Background: It has been described that an exposition of males to chemical substances may significantly impoverish quality and quantity of produced spermatozoa. The aim of our study was to test whether the polluted air in the Teplice district has negative effects on the quality of sperm of males living in this district., Methods and Results: 325 males 18-year-old living in the Teplice district and in the control district of Prachatice were tested. Samples were taken in 1992 and 1994, always at the end of winter and in autumn. According to WHO laboratory manual for investigation of the human sperm, basic parameters were determined: volume of the semen, pH, motility, number and morphology of spermatozoa. In selected groups of males the frequency of aneuploidia of spermatozoa was also examined. Examination of aneuploidia was done using three color fluorescence in situ hybridisation with satellite DNA proves specific for X, Z and 8 chromosomes. Logistic regression was used for the data analysis and Odd's Ratio was estimated (OR's). OR's was found for the morphology of spermatozoa (4.1 and 10.1 for medium and high exposition respectively), for the head morphology (6.1 and 4.1) and in the percentage of motile spermatozoa (9.8 and 3.5). More intensively exposed males had higher frequency of disomy in chromosomes X (p = 0.012), XY (p = 0.01), and Y (p < 0.001)., Conclusions: Bio-indicators of toxic and genetic impairment have shown lower quality of sperm in males in Teplice district.

A new centric fusion translocation, rob(16;20), was discovered in a phenotypically normal bull. C-banding revealed the dicentric nature of this centric fusion. This bull is a descendant of a German red pied bull and a Czechoslovakian red pied cow. Its mother and 26 half-brothers had normal karyotypes, indicating that this translocation arose "de novo".

The influence of silage juice feeding on the changes in 24 haematological and biochemical parameters in blood (blood serum) and urine of feeder bulls was evaluated in two comparative experiments in which no silage was included in feed rations. The supplement of 2 or 4 litres of juice per 100 kg of live weight represented a daily acidogenous stress of 0.79 to 1.93 moles of organic acids. In the first experiment this supplement was not proved to evoke any significant changes in the evaluated parameters. In the second experiment, in which the silage juices were enriched with sodium chloride, aqueous ammonia solution (26%) and molasses, a statistically significant increase in the level of beta-hydroxybutyric acid in blood and in the corresponding sum of ketone bodies (P less than 0.05 to 0.01) was observed in the experimental group of animals. The levels of sodium in serum were also increased (P less than 0.05). The average weight gains of bulls belonging to the experimental group were higher than those in the control groups.

The animals examined were 30 sows after eleven or more farrowings (pluriparous sows) and 26 primiparous sows. The pluriparous animals were 75.1 months old on the average and had farrowed an average of 13.23 litters. The mean age of the primiparous sows was 12.8 months. The blood sera of pluriparous sows had higher concentrations of total protein and urea and lower levels of 11-hydroxycorticosteroids (11-OHCS) and lower aspartate aminotransferase, alanine aminotransferase and alkaline phosphatase activities than those of primiparous sows. The pluriparous sows showed mild hypoglycaemia and hypocholesterolaemia. When insulin was injected i.m. in the dose of 1 I.U. per kg body mass, a decrease in blood serum glucose, total protein and potassium levels was recorded 60 min later in both pluriparous and primiparous sows; a significant rise in 11-OHCS concentration occurred only in the primiparous sows. It was concluded that the response of the hypothalamo-pituitary-adrenal axis in pluriparous sows to an insulin load is reduced. Studies of pluriparous sows contribute to a better understanding of the ontogenetic development of pigs throughout their postnatal development.

Circadian dynamics of 38 biochemical parameters of the blood, serum and urine of heifers was studied under the natural light conditions of a light (160-180 lux) and dark (10-40 lux) small cow house. In each house, eight parameters were subject to a significant influence during the day, six of them being the same in both houses: calcium, chlorides, urea and thyroxine of blood serum, glucose and haematocrit in the blood of heifers. The comparison of parameters in different groups showed that different light regimes exerted a significant influence on calcium, chlorides, total protein, cholesterol, vitamin A and thyroxine in serum, haematocrit and leucocyte count in blood, and sodium and urea in the urine of heifers. The effect of feeding regime on the dynamics of parameters was tested by the calculation of the percentual fluctuation of their average values in heifer blood and serum before feeding (samples taken at 6.30 hrs) and after feeding (sampled at 10 hrs). In the light house, six parameters were judged to be less stable and one unstable (urea in serum). In the dark house six parameters were less stable and four unstable (calcium, urea, copper and thyroxine in serum).

In farm production conditions two comparative trials lasting 58 and 128 days were carried out. Acidogenic exposure to acids of silage juices, silage and haylage (lactic acid, acetic and butyric acids) in feed ration ranged from 0.33 to 2.85 mol per 100 kg live weight. It was proved that it was possible to include 9 to 10 litres of silage juices in feed ration under the condition that the total daily intake would not exceed 1.94 mol of acids per 100 kg live weight. This supplement of silage juices increased milk production. Higher amount of acid (2.65 and 2.85 mol) resulted in acidogenic and ketogenic type of silage juice enriched feed ration, which caused a more rapid decrease of lactation curve in dairy cows fed this feed ration than in those in the control group.

Biochemical and hematological blood parameters were investigated in pigs experimentally infected with Ascaris suum. The most marked changes were recorded on day 5-8 post inoculation, during fever. In this period there was a marked decrease of concentrations of albumin, iron, thyroxin, A and E vitamins in blood serum, while compared with the control group the contents of A vitamin in liver did not change. Further, the concentrations of nonesterified fatty acid increased, as well as those of inorganic phosphorus and 11-hydroxycorticosteroids. Till day 13 PI the decrease of albumin and iron concentrations intensified and a recession of cholesterol serum binding capacity for iron was also recorded. Our results suggest that the acute phase of pig ascariasis is accompanied by changes in the host metabolism, generally corresponding to findings in fever states of different aetiology.