Synthesis and biological properties of new chimeric galanin analogue GAL(1-13)-[Ala10,11]ET-1(6-21)-NH2.

Several chimeric peptides consisting of the N-terminal fragment of galanin (GAL) and C-terminal fragments of other bioactive peptides (e.g. substance P, bradykinin, neuropeptide Y, mastoparan) have been synthesized and reported as high-affinity galanin receptor antagonists. Recently we have synthesized a new chimeric peptide, GAL(1-13)-[Ala(10,11)]ET-1(6-21)-NH(2), consisting of the N-terminal fragment of GAL and the C-terminal fragment of endothelin-1 (ET-1) analogue. This chimera was previously shown to be a moderate-affinity ligand to hypothalamic galanin receptors with a K(D) value of 205 nM. However, its biological action has been unknown so far. In our studies we characterized the biological properties of this new chimeric analogue, investigating its action on rat isolated gastric smooth muscles and influence on insulin secretion from rat isolated islets of Langerhans. Data acquired in the course of our studies suggest that analogue GAL(1-13)-[Ala(10,11)]ET-1(6-21)-NH(2) does not seem to be a potent galanin receptor antagonist in the gastrointestinal tract.

HPV and histological status of pelvic lymph node metastases in cervical cancer: a prospective study.

AIMS: The purpose of this prospective study was to describe the incidence and distribution pattern of human papillomavirus (HPV) DNA in intraoperative dissected lymph nodes and to relate this to the pathological confirmation of metastasis. METHODS: Samples of primary cervical cancer lesions and dissected lymph nodes were obtained from women undergoing surgical treatment. The presence of HPV DNA was detected by the polymerase chain reaction. RESULTS: Tissue from 79 tumours and 365 lymph nodes was analysed. Metastasis to the lymph nodes was found in 19 cases. Metastasis correlated with the volume of the primary lesion, the depth of cervical and vaginal invasion, and with invasion of the corpus. HPV DNA was found in 60 of the primary lesions and 31 of the lymph nodes. The presence of HPV DNA in the lymph nodes correlated with the volume of the primary lesion and vaginal invasion. CONCLUSIONS: The incidence of HPV DNA in lymph nodes is twice as high as that of histopathologically confirmed metastases. The risk of the presence of HPV DNA and histopathologically confirmed metastases in lymph nodes is related to certain features of the primary tumour.

[The use of PCR technique in the evaluation of the frequency of HPV infection in vulvar and cervical carcinoma]. / Zastosowanie techniki PCR do oceny czestosci wystepowania infekcji HPV w rakach sromu i szyjki macicy.

OBJECTIVES: In this study we estimated the frequency of human papillomavirus (HPV) infection in patients with vulvar and cervical carcinoma by polymerase chain reaction (PCR). We took into consideration the age of patients, the HPV infection status and frequency of HPV types. MATERIALS AND METHODS: Vulvar and cervical smears obtained from 88 patients with vulvar and cervical carcinoma, treated in 1997 in II Department of Obstetrics and Gynecology, Medical University in Gdansk were investigated for HPV DNA. We used the multiplex polymerase chain reaction (M-PCR) with four primer pairs: two primer pairs located in the E6, E7 open reading frame (ORF) for HPV6, 11, 16, 18, 31, 33, 35, 39, 45, 52b i 58 and primer par located in L1 ORF and type-specific primer for HPV16 (E6 ORF). We used restriction fragments length polimorphism PCR (RFLP-PCR) method for HPV typing. RESULTS: HPV DNA was detected in 3 (19%) of the 18 patients with vulvar carcinoma, of whom 3 (100%) had HPV16. In cervical carcinoma, DNA HPV identified in 50 (71%) of the 70, of whom 43 (86%) had HPV16, 3 had HPV31, 3 HPV33 and one was positive for HPV52-b. CONCLUSION: We did not find significant differences between the frequency of the HPV infection in different age groups of patients with cervical carcinoma. In vulvar cancer, we found the HPV infections significantly more often in group of patients in the age below 55.

[Frequency of appearance of antibodies to bovine serum albumin in children with diabetes type I]. / Czestosc wystepowania przeciwcial przeciwko albuminine wolowej (BSA-Ab) u dzieci ze swiezo wykryta cukrzyca typu 1.

It has been suggested that antibodies to bovine serum albumin may participate in the autoimmune process leading to the destruction of pancreatic islets. In the present study the frequency of antibodies to bovine serum albumin (BSA-Ab) in 45 children with newly diagnosed diabetes type 1, 32 children with diabetes lasting from 1-10 years and 65 healthy children was evaluated. BSA-Ab were determined by fluoroimmunometric method. The average value of fluorescence intensity 19 children with newly diagnosed diabetes type 1, who feeding mother's milk was 2037 x 10(3) +/- 898 x 10(3) impulse of fluorescence per minute (IMF) and was significant lower than the average value of fluorescence intensity of children with newly diagnosed diabetes type 1, who feeding artificiality. Antibodies to bovine serum albumin were found in 10% of children with newly diagnosed diabetes type 1, who feeding at the first six months of their life mother's milk, in 42% of children with newly diagnosed diabetes type 1, who feeding artificially and 3% of healthy children.

[Large-scale screening for early diagnosis of gestational diabetes mellitus (GDM)]. / Wczesne rozpoznawanie cukrzycy ciazowej w populacyjnych badaniach przesiewowych.

The aim of study was to evaluate incidence of GDM in Poland. All 1500 pregnant women between 24-28 week's gestation consulted in 4 centers were offered a 50 g oral glucose test (screening test). Capillary blood glucose was measured at 60 min after glucose ingestion. When blood glucose > 140 mg/dl, 75 g OGTT according to WHO criteria was performed. 241 women have abnormal screening test and in 181 cases blood glucose were at range 140-160 mg/dl, in 39 at range 160-180 and 21 were > 180 mg/dl. Only 14 women in the first group (140-160 mg/dl) have diagnosed GDM (7.7%). In second group 24 pregnant women have GDM (61.5%). Overall GDM incidence is shown to be 3.7% (57 women). The mean age for the GDM was 28.8 +/- 0.9 years compared with 26.4 +/- 0.4 years (p < 0.05) uncomplicated pregnancy.

[Changes in levels of human placenta lactogen (hPL), progesterone, and estriol in blood serum and estrogens in urine during gestational diabetes mellitus]. / Zmiany poziomu laktogenu lozyskowego (hPL), progesteronu, estriolu w surowicy oraz estrogenów w moczu w ciazy powiklanej cukrzyca ciezarnych (GDM).

The aim of the study was estimation of endocrinological function of placenta in pregnancy complicated by GDM. The study were performed on a group 13 women with GDM and 14 women in normal pregnancy. All women with GDM were treat by diet and intensive insulinotherapy with self monitoring levels of glucose. In women with GDM level of fructosamine and HbAlc were significant higher but in normal range. In 28, 36, 37, 38, 39 week of pregnancy were determined levels of hPL, oestriol and progesterone in serum and daily excretion of oestrogens in urine. From 36 week of pregnancy levels of hPL, oestriol and progesterone were significant lower in women with GDM. In 39 week of pregnancy level of hPL was 9.34 micrograms/ml vs. 12.71 micrograms/ml, p < 0.01, oestriol was 495.77 nmol/l vs. 681.14 nmol/l, p < 0.01 and progesterone was 25.12 ng/ml vs. 34.52 ng/ml, p < 0.01 respectively in women with GDM and in normal pregnancy.

[Changes in levels of chorionic gonadotrophin (hCG) and its subunit alpha and beta in pregnancy complicated by diabetes (GDM)]. / Zmiany poziomu gonadotropiny kosmówkowej (hCG) oraz jej podjednostek alfa i beta w ciazy powiklanej cukrzyca ciezarnych (GDM).

The aim of the study was estimation of endocrinological function of placenta in pregnancy complicated by GDM. The study were performed on a group 13 women with GDM and 14 women in normal pregnancy. All women with GDM were treat by diet and intensive insulinotherapy with self monitoring levels of glucose. In women with GDM level of fructosamine and HbAlc were significant higher but in normal range. In 28 and 36 week of pregnancy were determined levels of hCG, alpha hCG, beta hCG, in serum. Level of hCG in control group and in women with GDM were respectively 97.29 U/ml vs. 29.29 U/ml, p < 0.01 in 28 week of pregnancy and 77.23 U/ml vs. 37.93 U/ml, p < 0.05 in 36 week. Level of alpha hCG was lower and beta hCG was higher in group with GDM.

[Transplantation of free beta islet cells as a method leading to normalization of glucose tolerance in rats with experimental diabetes]. / Transplantacja wolnych komórek beta wysp trzustki jako metoda wyrównania zaburzen tolerancji glukozy u szczurów z cukrzyca doswiadczalna.

The aim of present study was to investigate the effect of free beta cells transplantation on carbohydrate tolerance in rats with streptozotocin-induced diabetes. Free beta cells were isolated by mechanical disruption of isolated pancreatic islets in Ca and Mg free medium. 1500 and 3000 isolated islets were used for the isolation of 500 x 10(3)-1.099 x 10(3) and 1.398 x 10(3)-2.098 x 10(3) free cells, respectively. Between 78% and 98% of all isolated cells were the variable cells. After isogenic transplantation of free beta cells, blood glucose concentration in all animals was normalized within 14 days. The rate of glycemia normalization was related to the amount of viable free cells. The glucose assimilation coefficient after intravenous glucose administration in normal rats, diabetic rats and diabetic rats after transplantation of free cells was: 2.98 x 10(-2); 0.68 x 10(-2) and 2.04 x 10(-2) mg/dl/min, respectively. Allogenic transplantation of free beta cells caused only a transient decrease in blood glucose level.

Protective effect of allopurinol, alpha-tocopherol and chlorpromazine on rat pancreatic islets stored prior to transplantation.

Iso-transplantation of 800 isolated rat pancreatic islets into the portal vein of diabetic rats was performed. Isolated islets were kept in Hanks buffer for 12 hours prior to transplantation. Part of donors was pretreated with allopurinol, alpha-tocopherol and chlorpromazine. Transplantation of islets isolated from nonpretreated rats did not cause any significant changes in plasma glucose concentration of recipients, while transplantation of islets isolated from donors after pretreatment with the tested substances brought glucose level back to normal 3 days after transplantation. Our results indicate that the combination of free radical scavengers, antioxidants and membrane stabilizing drugs may be used to increase the effectiveness of islet transplantation in humans.

Binding of 3H-norepinephrine to receptors of pancreatic islets isolated from neonatal and adult rats.

Binding of 3H-norepinephrine to receptors and insulin release from pancreatic islets isolated from newborns and adult rats were investigated. In the presence of norepinephrine at the concentration of 2.4 x 10(-8) mol/l adults islets bound 0.37 +/- 0.11 fmol of norepinephrine per micrograms of islet protein while neonatal islets bound 0.25 +/- 0.08 fmol/micrograms. When norepinephrine concentration was raised to 10(-5) mol/l the amount of bound norepinephrine increased up to 22.66 +/- 8.7 fmol/micrograms and 14.20 +/- 5.36 fmol/micrograms for both examined groups, respectively. Addition of 5 x 10(-4) mol/l of phentolamine to the incubation medium induced 74% and 70% decrease of bound norepinephrine to adult and neonatal islets, respectively. At similar range of concentrations both norepinephrine and phentolamine modified the rate of insulin secretion and caused changes in binding of 3H-norepinephrine. There were no differences between the binding of norepinephrine to neonatal and adult islets. This may suggest that adrenergic system may play greater role in the regulation of insulin release from neonatal than from adult islets.

[Enalapril in patients with diabetes mellitus type I with microalbuminuria without hypertension]. / Enalapryl u chorych na cukrzyce typu I z mikroalbuminuria bez nadcisnienia tetniczego.

The studies included 18 normotensive patients with diabetes mellitus type I (mean age 29 years) and constant microalbuminuria (UAE-30 - 300 mg/24 hours). Group A consisted of 10 patients treated with enalapril, and group B--10 patients given placebo. Glomerular filtration rate, ERPF, and UAE were measured before and after 6 months of therapy. UAE decreased significantly in patients of group A (p = 0.02) after 6 months while evident proteinuria was seen in two patients of group B. Arterial blood pressure dropped in patients of group A (131/84 vs 122/78 mm Hg), and increased significantly in patients of group B (126 +/- 8 vs 136 +/- 15 mm Hg; p < 0.05). Blood flow through kidneys improved (p = 0.02) and renal vascular resistance decreased (p = 0.02) in patients of group A. The obtained results suggest that enalapril may prevent diabetic nephropathy in diabetics with constant microalbuminuria.

Lack of effect of high-dose biosynthetic human C-peptide on pancreatic hormone release in normal subjects.

We studied the effect of high doses of biosynthetic human C-peptide on pancreatic hormone secretion in response to oral (75 g) and intravenous [( IV] 0.33 g/kg of D50%) glucose on normal volunteers. The infusion of human C-peptide at a rate of 360 ng/kg/min body weight, increased the plasma C-peptide concentration from a basal level of 0.32 +/- 0.04 pmol/mL to 38.5 +/- 1.8 pmol/ml. Overall, C-peptide had no significant effect on the serum levels of glucose, insulin, proinsulin, glucagon, and pancreatic polypeptide, either under basal conditions or following IV and oral glucose administration. However, small decreases in glucose and insulin concentrations that were not statistically significant were seen during the first hour after C-peptide infusion. The results of the present studies are therefore consistent with the conclusion that even supraphysiologic plasma concentrations of infused C-peptide do not affect basal insulin secretion or overall insulin secretory responses to oral or IV glucose. However, we cannot definitively exclude a small reduction in insulin secretion in the first hour after oral glucose ingestion.

Expression of Ly-6C by T lymphocytes of NOD mice after CD3-complex stimulation. Identification of activated cells during insulitis of prediabetic mice.

Ly-6C is a differentiation antigen that distinguishes T-lymphocyte subsets. In concordance with previous results, splenocytes from NOD mice do not express the epitope recognized by anti-Ly-6C monoclonal antibodies (MoAbs), including MoAb HK1.4 in this study, and cannot be stimulated to proliferate in response to HK1.4. However, when splenocytes from NOD mice were stimulated in vitro with the anti-CD3 MoAb 145-2C11, T lymphocytes expressing Ly-6C were detected after 48 h of stimulation, with as many as 25% of lymphocytes expressing this antigen with prolonged passage in culture. Most of the cells expressing Ly-6C were Thy-1.2+, CD4+, and CD8- and proliferated after stimulation with HK1.4. To further understand the failure of NOD splenocytes to express Ly-6C, freshly isolated cells were stimulated with alpha/beta-interferon (IFN-alpha/beta) and IFN-gamma. Although these lymphokines induced expression of Ly-6A and Ly-6C in splenocytes from C57BL/6J mice and Ly-6A in NOD cells, Ly-6C was not induced on NOD cells. Because Ly-6C expression on splenocytes was a marker of activation via the CD3 T-lymphocyte receptor complex, we also examined expression of Ly-6C on T lymphocytes within islets showing insulitis in vivo. Lymphocytes that were Ly-6C+ were identified within islets on histological sections of pancreas, whereas Ly-6C+ cells in the spleen from the same mouse could not be detected. Our findings imply functional abnormality in expression of Ly-6C in NOD mice.(ABSTRACT TRUNCATED AT 250 WORDS)

[Derivatives of 2-mercaptobenzenesulphonylamide. IV. Synthesis and properties of some hypoglycemic agents derived from N-(4-chloro-2-mercapto-5-methylbenzenesulphonyl)urea]. / Pochodne 2-merkaptobenzenosulfonamidu. IV. Syntezy i wlasciwosci hipoglikemizujace niektórych pochodnych N-(4-chloro-2-merkapto-5-metylobenzenosulfonylo)mocznika.

Some new S- and N'-substituted derivatives of N-(4-chloro-2-mercapto-5-methylbenzenesulphonyl)urea were obtained. Their structures were confirmed by elemental analysis, and by IR and NMR spectra. Schemes of the reactions, their yields and some physico-chemical and pharmacological properties of the compounds are also given. N-(4-chloro-5-methyl-2-methylthiobenzenesulphonyl)-N'-R-ureas (I, II, IV) were screened for hypoglycemic activity in rats, and the results are compared to effect of tolbutamide.

[Clinical examination of diabetic patients treated with monocomponent insulin manufactured by MP Polfa]. / Badania kliniczne chorych na cukrzyce leczonych insulina jednoszczytowa MP Polfa.

An effect of replacing conventional forms of insulin by the monocomponent insulin manufactured by "Polfa" was studied in the group of 22 diabetics. The patients were followed up for 12 months. An effect of monocomponent insulin on daily requirement of insulin, levels of anti-insulin, monocomponent and pancreatic peptide antibodies, compensation of diabetes mellitus, and lipodystrophy were investigated. New insulin preparation decreased anti-insulin and pancreatic peptide antibodies level and markedly diminished lipodystrophy. However, daily insulin requirement, degree of diabetes mellitus compensation, and anti-proinsulin antibodies level remained unchanged.