Therapeutic antitumor potential of endoglin-based DNA vaccine combined with immunomodulatory agents.

Therapy targeting tumor blood vessels ought to inhibit tumor growth. However, tumors become refractory to antiangiogenic drugs. Therefore, therapeutic solutions should be sought to address cellular resistance to antiangiogenic therapy. In this regard, reversal of the proangiogenic and immunosuppressive phenotype of cancer cells, and the shift of the tumor microenvironment towards more antiangiogenic and immune-stimulating phenotype may hold some promise. In our study, we sought to validate the effects of a combination therapy aimed at reducing tumor blood vessels, coupled with the abrogation of the immunosuppressive state. To achieve this, we developed an oral DNA vaccine against endoglin. This antigen was carried by an attenuated Salmonella Typhimurium and applied before or after tumor cell inoculation into immunocompetent mice. Our results show that this DNA vaccine effectively inhibited tumor growth, in both the prophylactic and therapeutic settings. It also activated both specific and nonspecific immune responses in immunized mice. Activated cytotoxic T-lymphocytes were directed specifically against endothelial and tumor cells overexpressing endoglin. The DNA vaccine inhibited angiogenesis but did not affect wound healing. In combination with interleukin-12-mediated gene therapy, or with cyclophosphamide administration, the DNA vaccine resulted in reduced microvessel density and lowered the level of Treg lymphocytes in the experimental tumors. This effectively inhibited tumor growth and prolonged survival of the treated animals. Polarization of tumor milieu, from proangiogenic and immunosuppressive, towards an immunostimulatory and antiangiogenic profile represents a promising avenue in anticancer therapy.

Overweight and obesity among adults in Poland, 1983-2005.

INTRODUCTION: Overweight and obesity are causes for numerous non-communicable chronic diseases. In recent years their incidence have become a major epidemiological problem both in Poland and many other countries. PURPOSE: Evaluation of the current situation of the overweight and obesity prevalence among adults in Poland and changes in this regard in the years 1983-2005. MATERIAL AND METHODS: Review of the countrywide research and research in selected towns and regions on overweight and obesity occurrence conducted in Poland over the last 20 years. RESULTS: On the basis of national data for Poland, the incidence of overweight is estimated to be 39-40% among men and 28-29% among women, while the percentage of obese men and women is 16-21% and 19-22% respectively. The incidence of overweight and obesity differs with respect to gender, age, environment, the region of the country and social status. Excessive body weight is often observed in patients treated in hospitals. Abdominal obesity is very common and according to the ATP III guidelines, affects 16-28% of men and 35-40% of women, whereas under more strict IDF criteria it involves almost 40% of adult males and almost a half of adult females. CONCLUSIONS: Results from numerous surveys conducted in the last few years indicate growth of the incidence of overweight and obesity, especially among men. The prevalence of overweight and obesity in Poland is comparable with most European countries.

Comparison of the toxicity and distribution of cadmium and lead in plant cells.

The toxicity of heavy metals (Cd, Zn, and Pb) was assessed by in vivo observations of their effect on cytoplasmic streaming in Allium cepa L. bulb scale epidermal cells. On the basis of our results, the order of toxicity of the studied cations is Zn < Pb << Cd. The difference in toxicity between cadmium and lead was found to be very large. When cytoplasmic streaming was assessed, this difference was threefold. When the total content of cadmium and lead (determined by inductively coupled plasma mass spectrometry) was the criterion, the difference in toxicity was 15-fold. Fractionation of the tissue and enzymatic digestion of the cells revealed that the largest proportion of cadmium was located in the cell walls (56%), whereas almost all of the lead (97.6%) was accumulated in an insoluble form. The speciation of water-soluble Pb and Cd fractions is discussed on the basis of analysis by capillary zone electrophoresis interfaced with inductively coupled plasma mass spectrometry of water extracts from epidermal cells. Lead and cadmium appeared to be bound mainly to salts, which explains their toxicity. Cadmium was complexed (detoxified) by organic acids, while thiols were the metal-complexing species for lead. Histidine formed complexes with both cadmium and lead. Ultrastructural analyses showed that lead was encapsulated in small vesicles in the cytoplasm. Fluorescence studies of the endoplasmic reticulum (ER) revealed that it underwent extensive fragmentation under the influence of lead, with numerous ER vesicles appearing in the cells. In other words, the lead deposits in the cytoplasm were contained in vesicles arising from fragmentation of the ER. These observations indicate that epidermal cells have a rapid and effective mechanism for detoxifying lead involving the ER, and this may be one of the mechanisms accounting for the lower toxicity of lead in comparison with cadmium. The suitability of Allium cepa bulb scale epidermal cells for use in ecotoxicological studies is also discussed. Step-by-step directions for this test are given.

Counteraction against obesity--is it possible?

The obesity epidemic is one of the most serious public health problems across many countries. In Poland more than half of the adult population has excessive body weight, while approx. 20% are obese. 15-20% of children and adolescents suffer from excessive body weight, while 4% of them are obese. Moreover, the number of overweight or obese children is growing alarmingly. Obesity can lead to many serious health consequences. Though the most serious disorders are cardiovascular diseases, diabetes type 2 and some cancers. In the nearest future diseases related to obesity will probably become the main cause of death in many countries. This may lead to shorter average life expectancy. The treatment costs of obesity and related diseases are constantly increasing. The most important preventive measure aiming at curbing the effects of obesity involves lifestyle change, including a change in diet and physical activity. The best results should be obtained by multifaceted programmes, which cover activities aiming at the improvement of both diet and physical activity. Due to the spread of the obesity epidemic, the countries of WHO European Region signed the European Charter on Counteracting Obesity, in which they declared their commitment to combat obesity. Activities aiming at combating obesity in Poland should be closely connected with the implementation of the National Programme for the Prevention of Overweight, Obesity and NonCommunicable Diseases through Diet and Improved Physical Activity, which will be implemented in 2007-2016.

Guidelines in the medical treatment of Helicobacter pylori infection.

Scientific evidence based on controlled clinical research confirm substantial benefits resulting from the eradication of H. pylori infection in such pathologies of the alimentary tract as: gastric peptic and duodenal ulcer (active or confirmed in the future and ulcer disease complications), MALT (Mucosa Associated Limphoid Tissue) lymphoma, atrophic gastritis, past stomach resection, gastric cancer in the family. The above group of indications is strongly recommended for eradicative treatment. During the last several years there have been many guidelines made by international and national specialist groups. "Test and treat" strategy of undiagnosed dyspepsia treatment is based on possibility to carry out non-invasive tests confirming H. pylori infection. First symptoms of dyspepsia in people over 45 years of age constitute recommendation for endoscopy, as well as symptoms assumed to be "alarming" (loss of weight, anaemia, bloody vomiting, tarry stool, dysphagia) regardless of patient age. An individual approach to eradication is proposed in gastroesophageal reflux disease, and use of non-steroid anti-inflammatory drugs. Antibacterial activity towards H. pylori is shown by many antibiotics (amoxicillin, macrolides, tetracyclines) and some other chemotherapeutic agents (nitroimidazoles) and bismuth. PPIs are recommended, because through increase of pH in stomach they create conditions to act for antibiotics. During the stage of first line triple therapy, it is advised to apply PPI and two antibacterial medicines at the same time (PPI + amoxicillin+metronidazole or clarithromycin). Such therapeutic action ensures achievement of eradication of H. pylori infection in 80-90% of cases. In case of lack of treatment efficiency in the first-line therapy, 7-14 day treatment may be repeated using triple therapies (PPI + 2 antibiotics) substituting the antibiotic with the metronidazole or tetracycline, or quadruple therapies (PPI + bismuth citrate + 2 antibiotics). Side effects during eradicative treatments occur quite rarely (from 15 to 30%).

Nutritional status of patients hospitalised in Poland.

AIM: Screening and extended assessment of the nutritional status of patients on admission and on discharge from hospital were carried out. DESCRIPTION: The studies were carried out in four teaching hospitals, four provincial hospitals and four county hospitals in Poland. SUBJECTS: Screening examinations were carried out for 3310 randomly selected patients (every 10th patient admitted to hospital, including 1916 female cases aged from 16 to 92 y and 1394 male patients aged from 16 to 100 y). Extended examinations were carried out on 210 patients aged from 16 to 87 y (including 122 female and 88 male). MAIN ASSESSMENT PARAMETERS: Anthropometric (height, weight, body mass index (BMI), waist-to-hip ratio (WHR), arm circumference) and biochemical indices (erythrocyte count, haemoglobin concentration, white blood cell count, blood lymphocyte count and serum albumin serum concentration). The extended examinations included determination of antioxidant vitamins (A, C, E), vitamin B(12) and folic acid. RESULTS: On admission to hospital, 10.43% of the patients surveyed had a BMI below 20 kg/m(2), in 20.74% of patients serum albumin concentration was below 3.5 g/dl, indicating possible protein energy malnutrition. In addition, 21.02% had lymphocyte count below 1.5 x 10(3)/mm(3). During hospitalisation, deterioration in the nutritional status of the patient population occurred. On discharge from hospital, the percentage of patients with BMI < 20 kg/m(2) increased to 11.21% and the percentage with low blood albumin (<3.5 g/dl) increased to 28.57%. On admission, vitamin C deficiency was present in 51.8% of patients, folic acid deficiency in 32%, vitamin E deficiency in 10%, vitamin B(12) deficiency in 6.8% and vitamin A deficiency in 1.4%. Vitamin deficiencies were present equally in malnourished, overweight and obese patients. CONCLUSIONS: In patients admitted to hospitals in Poland, malnutrition risk demonstrated by BMI was observed in 10.43% of patients. On the basis of biochemical indices, increased nutritional risk was demonstrated in 21% of patients. Vitamin malnutrition was seen in the majority of patients. A significant correlation between weight, BMI, arm circumference, blood lymphocyte count and the number of days spent in hospital was observed. SPONSORSHIP: The Committee of Scientific Research and the Ministry of Health-PBZ 012-14.

Atmospheric pressure chemical ionization mass spectrometric and visible spectrophotometric studies of copper(I) and copper(II) complexes with 2-(5-bromo-2-pyridylazo)-5-diethylaminophenol.

Complexes of copper(II) with 2-(5-bromo-2-pyridylazo)-5-diethylaminophenol (5-Br-PADAP) formed in aqueous methanol in a wide range of pH (from acidic to alkaline) as well as copper(I)-5-Br-PADAP species formed in methanolic solutions were investigated by spectrophotometry and mass spectrometry. Pseudomolecular and fragment ions created in the atmospheric pressure chemical ionization (APCI) source confirmed the molecular masses of the complexes existing in the solvents and their structures. The structure of the Cu(II) complex with 5-Br-PADAP formed in acidic medium was proposed as CuR(R - H) (where R is the undissociated molecule of the reagent). The binding sites of the two bound reagent molecules were different: in one of them the oxygen atom of the dissociated phenolic group and the nitrogen atom from the azo (-N=N-) group took part in complex formation, whereas in the other only nitrogen atoms from the pyridyl ring and azo group were involved. The complex was stable and could not be reduced to Cu(I) species by use of standard reducing agents (ascorbic acid, hydroxylamine). In alkaline solutions the complex tended to polymerize and precipitated in media containing less than 80% of methanol. The copper(I)-5-Br-PADAP complex was extremely unstable and could be obtained (as a mixture with Cu(II) species) in media free of water or oxygen. For this complex, CuR(2) was proposed as the most probable structure. According to this proposal copper(I) reacted exclusively with nitrogen-containing binding sites and the undissociated phenolic group was not engaged in complex formation. In this system Cu(I)/Cu(II) electron transfer is very rapid, accelerated by a polar environment, e.g. in the presence of water molecules or dissolved oxygen.

Tobacco smoking and vitamin C concentration in gastric juice in healthy subjects and patients with Helicobacter pylori infection.

Low gastric juice total vitamin C concentration in the presence of Helicobacter pylori infection probably plays a role in gastric carcinogenesis. Vitamin C plays a role in the neutralization of various pathogenic factors connected with H. pylori infection, including the destruction of free radicals, which damage tissues and cell DNA, and inhibition of the formation of N-nitroso compounds, which have a strong carcinogenic activity. The aim of the study was to determine whether tobacco smoking had any effect on gastric juice vitamin C concentration in healthy subjects and in patients infected with H. pylori. Eighty-six patients with dyspeptic symptoms undergoing routine endoscopy entered the study after giving informed consent. In all patients plasma and gastric juice total vitamin C levels were measured by a spectrophotometric method. They were entered into four groups: group I (controls) - H. pylori-negative non-smokers (n = 17), group II - H. pylori-negative smokers (n = 16), group III - non-smokers with H. pylori infection (n = 21), and group IV - H. pylori-infected smokers (n = 32). In the control group (I) the mean gastric juice total vitamin C concentration was 17.1 microg/ml (range 5.3-40.0 microg/ml), which was significantly higher (P < 0.05) than in group II (12.6 microg/ml, range 5.1-21.0 microg/ml), group III (5.8 microg/ml range 2.1-13.7 microg/ml) and group IV (3.9 microg/ml, range 1.1-10.6 microg/ml) (P < 0.001). Statistically significant differences also were noted between groups II and III (P < 0.01) and groups II and IV (P < 0.001) and between groups III and IV (P < 0.05). These results demonstrate that the concentration of vitamin C in gastric juice is significantly lower in smokers than in non-smokers. This was observed in healthy subjects as well as H. pylori-infected patients. This phenomenon may be one of the mechanisms whereby smoking contributes to the production of gastric lesions, impairs healing of peptic ulcers and also increases the recurrence rate of peptic ulcers in cases with H. pylori infection.

[Interactions between food and drugs. 2. Disturbances of drug metabolism and synergistic mode of action]. / Interakcje miedzy zywnoscia a lekami. 2. Zaburzenia metabolizmu leków i dzialania synergiczne.

The paper overviews food-drug interactions. The part 2 reviews the interactions of foods with the drugs which occur at the level of metabolism. Concomitant intake with food increases or decreases the concentrations of many agents in humans. The effect seems to be mediated mainly by suppression of the cytochrome P450 in the liver and in the small intestine wall. This results in a diminished first pass metabolism with higher bioavailability and increased maximal concentrations of substrates of this enzyme. To avoid the interactions many drugs should not be taken with the meal and citrus juices, particularly grapefruit juice.

[Interactions between food and drugs. 1. Malabsorption]. / Interakcje miedzy zywnoscia a lekami. 1. Zaburzenia wchlaniania.

The paper overviews food-drug interactions. The part 1 reviews the interactions of foods with the many different agents which occur at the level of absorption, considering the mechanisms involved. The food-drug interactions can cause an increase, decrease, or delay in the bioavailability of the drugs. There is an aspect of therapy which may have an important clinical repercussion, which it must not be forgotten.

Particular fractions of microproteinuria in patients with stabile angina pectoris and without a clinical nephropathy.

The determination of microalbuminuria is a valuable method in the diagnosis of renal and vascular diabetes or hypertension complications. Recently, microalbuminuria appeared to be the predictor of coronary heart diseases (CHD). The presented study comprised 26 patients with stable angina pectoris (AP) and 27 healthy volunteers. We simultaneously evaluated microproteinuria during the first morning and afternoon miction and the 24-h blood pressure. Amongst patients with AP all urine protein concentrations were increased (results in g/mol creatinine): alpha-1-microglobulin (1.04 + 0.13 vs. 0.47 + 0.05, p < 0.001) albumin (0.95 + 0.15 vs. 0.61 + 0.05, p < 0.05) and IgG (1.00 + 0.17 vs. 0.55 + 0.05, p < 0.01) were higher, in comparison to control group values. Indices for diurnal blood pressure rhythm were significantly lower in the AP group for both systolic (1.07 + 0.01 vs. 1.14 + 0.01 p < 0.001) and diastolic (1.09 + 0.02; vs. 1.21 + 0.03 p < 0.01) pressures. A physiological increase of albumin from the afternoon sample was only observed in the control group. Thus, our AP patients demonstrated signs of subclinical nephropathy in both the proximal tubuli and glomeruli.

[The selectivity, polarity and characteristic features of autistic and syntonic schizophrenia]. / Selektywnosc, biegunowosc i cechy charakterystyczne schizofrenii autystycznej i syntonicznej.

In previous papers (3, 4, 5) the author presented a hypothesis concerning important features of clinical picture of paranoid schizophrenia, that is selectivity and polarity (bipolarity). Numerous research works aimed at closer recognition of some differences of clinical picture of paranoid schizophrenia have been conducted under the author's direction. The formation of certain psychopathological proportions discerned by the author has been researched with intentionally elaborated scales. The research results have led to distinguishing autistic and syntonic schizophrenia.

Reversed-phase liquid chromatographic determination of uranium based on its ternary complex with fluoride and 2-(5-bromo-2-pyridylazo)-5-diethylaminophenol. New chromatographic probe for the detection of fluoride on C18 stationary phase surface.

Optimum conditions for the reversed-phase liquid chromatographic separation of uranium as U(VI)-F(-)-(5-Br-PADAP) [5-Br-PADAP is 2-(5-bromo-2-pyridylazo)-5-diethylaminophenol] ternary complex on the end-capped C18 column were evaluated. The developed sensitive, selective and versatile method allows to determine uranium in the wide concentration range 0.2-120.0 mug ml(-1) (detection limit, DL, is 0.15 mug ml(-1) for 20 mul loop). Solvent mixture, acetonitrile+water (65+35, v/v) containing fluoride in concentration 3x10(-3) mol l(-1) (pH 5.5) was used as eluent. Double action of fluoride present in eluent - as stabilising agent of the ternary system and modifier of the stationary phase was evaluated. In order to prove appearance of the modifying effect of fluoride on the stationary phase the new chromatographic probe - system Zr(IV)-(5-Br-PADAP)/Zr(IV)-F(-)-(5-Br-PADAP) was introduced as a tool for the detection of F(-) presence on the surface.

Sensitive reversed-phase liquid chromatographic determination of fluoride based on its ternary systems with zirconium(IV) or hafnium(IV) and 2-(5-bromo-2-pyridylazo)-5-diethylaminophenol.

Optimum conditions for the direct reversed-phase LC determination of fluoride based on the ternary M-(F-)-(5-Br-PADAP) complexes [M = ZrIV or HfIV and 5-Br-PADAP = 2-(5-bromo-2-pyridylazo)-5-diethylaminophenol] were evaluated. Chromatographic separation was performed with C18 end-capped column with an eluent consisting of acetonitrile-water (85 + 15 v/v) mixture of pH 4.0 +/- 0.3 (flow rate 1 ml min-1), and the eluate was monitored spectrophotometrically at lambda max = 585 nm. The calibration curves were linear over a wide range of fluoride concentrations: from 1 to 110 and 150 ng ml-1 for the ZrIV-(F-)-(5-Br-PADAP) and HfIV-(F-)-(5-Br-PADAP) systems, respectively (using a 20 microliters loop). Under such conditions the detection limits were 0.8 and 0.7 ng ml-1, respectively, and the quantification limit is 1.0 ng ml-1 for both methods. When a 100 microliters loop was used, the limits of both detection and quantification in the method based on the zirconium system were 0.2 ng ml-1. Using the proposed method, fluoride was determined directly in tap water, saliva and an anti-cancer agent for prostatic cancer (Leuprolid).

Effects of high dose vitamin C treatment on Helicobacter pylori infection and total vitamin C concentration in gastric juice.

Low gastric juice total vitamin C concentration in the presence of Helicobacter pylori (H. pylori) infection probably plays a role in gastric carcinogenesis. In vitro vitamin C has been shown to inhibit the growth of H. pylori. The aims of this study were to determine the effect of high dose vitamin C administration on H. pylori infection and on gastric juice total vitamin C concentration in patients with H. pylori related chronic gastritis. Sixty patients with dyspeptic symptoms and proven chronic gastritis and H. pylori infection, who were undergoing routine endoscopy, entered the study after giving informed consent. They were randomly coded into two treatment groups. Group 1 (controls, n = 28) were treated with antacids for 4 weeks and Group 2 (n = 32) received vitamin C 5g daily also for 4 weeks. Nine patients did not complete the study and were excluded. Plasma and gastric juice total vitamin C levels were measured at baseline, at the end of 4 weeks treatment and again 4 weeks after treatment cessation. In the control group H. pylori infection remained unchanged in all 24 patients throughout as did the mean gastric juice total vitamin C concentration. However, in the vitamin C treated group eight of 27 patients (30%) who completed the treatment course the H. pylori infection was eradicated (P = 0.01). In these patients the mean gastric juice total vitamin C concentration rose significantly from 7.2 +/- 1.6 micrograms/ml after 4 weeks treatment (P < M 0.001) and 19.8 micrograms/ml 4 weeks after treatment was discontinued (P < 0.001). In the remaining 19 patients with persistent H. pylori infection, the mean gastric juice total vitamin C concentration rose less than in those with successful H. pylori eradication; 6.3 +/- 1.7 micrograms/ml before treatment, 10.8 +/- 1.5 micrograms/ml after 4 weeks treatment (P < 0.05) and a return to pre-treatment levels (7.1 +/- 2.7 micrograms/ml) 4 weeks after vitamin C intake stopped. There were no side effects of vitamin C treatment. This study has shown that 4 weeks daily high dose vitamin C treatment in H. pylori infected patients with chronic gastritis resulted in apparent H. pylori eradication in 30% of those treated. In those patients there was also a highly significant rise in gastric juice total vitamin C concentration which persisted for at least 4 weeks after the treatment ceased. A significant, though less marked, gastric juice total vitamin C concentration increase was observed during vitamin C treatment even in subjects with persistent H. pylori infection, though this was not maintained after treatment ended. The mechanism whereby vitamin C treatment appeared to result in H. pylori eradication is unclear. Further confirmatory studies are indicated.

[A case of chronic calcifying pancreatitis in a 19 year old man]. / Przypadek przewleklego wapniejacego zapalenia trzustki u 19-letniego mezczyzny.

We present an advanced case form of chronic calcifying pancreatitis (ch.c.p.) of 19's age man, whose first clinical symptoms appeared when he was 5 years old. For the rest 14 years his complains were diagnosed as vegetative neurosis. The patient was admitted in to the Department in 4th day of severe abdominal pain. High increase of serum and urine amylase activity, changes in USG and CT examinations (20 mm wide pancreatic duct with numerous calcified stones) were find. Acute pancreatitis in the course of ch.c.p. was diagnosed. After typical treatment abdominal pain disappeared and results of laboratory tests achieved a normal range. During 1.5 year follow up (diet, pancreatic enzymes) substitution abdominal pain did not appear and the body weight increased 8 kg.

[A case of ascites in the course of acute pancreatitis]. / Przypadek wodobrzusza w przebiegu ostrego zapalenia trzustki.

The case of acute alcoholic pancreatitis, complicated by internal pancreatic fistula is being reported. The general patients symptom in initial illness period was ascites in peritoneal cavity. Biochemical investigation of ascites has shown elevated activity of amylase and protein which is typical for pancreatic ascites. The late diagnosis of ascites cause (in 3rd week of the illness) and complications decreased efficacy of the treatment used (parenteral nutrition and somatostatin). Thus worse the course of the illness.