Effect of low temperature cultivation on insulin secretory of human pancreatic islets.

The experiment compared the physiological function (insulin secretory capacity) and membrane integrity of human adult pancreatic islets incubated in culture at 37°C and 24°C. Pancreatic tissue was digested with Collagenase XI, using a non-automated method. Cultures were incubated at 37°C and 24°C. Secretory capacity of the islets is determined by measuring of the stimulation index (SI) on the 1st, 3rd and 7th day of cultivation. Membrane integrity of the islets was determined by dithizone staining. Both groups of examined cultures show a slight increase in SI during the incubation. However islets incubated at 24°C show higher SI values than those incubated at 37°C on the 1st, 3rd and 7th day of incubation. And on the first day of incubation, this difference was statistically significant (p <0.05). Islets incubated at 37°C showed preservation of membrane integrity, the islets are regular spherical shape, while those incubated at 24°C lose such an organization. During the seven-day cultivation, islets incubated at a standard temperature of 37°C show less preserve physiological functions in relation to cultures incubated at 24°C, but islets incubated at 37°C show more regular morphological forms.

A novel Notch3 Gly89Cys mutation in a Serbian CADASIL family.

Cerebral autosomal dominant arteriopathy with subcortical infarcts and leukoencephalopathy (CADASIL) is the most common heritable cause of stroke and vascular dementia in adults. We present a family from Serbia presenting with stroke and depression in the lack of vascular risk factors, with brain MRI indicating CADASIL. A novel NOTCH3 Gly89Cys mutation was located in exon 3. This report illustrates that in the setting of a positive family history with typical clinical and MRI features, even with an atypical form of pedigree, a high suspicion of CADASIL should lead to genetic testing.

[Evaluation of pulmonary and metabolic function in classic and laparoscopic cholecystectomy in patients with ASA I and ASA II scores].

This is a prospective study of patients treated at the Center for Urgent Surgery, Clinical Center of Serbia. The patients were divided into two groups; i.e., the controls consisted of 30 subjects, who underwent conventional cholecystectomy, and studied group with 30 patients who had laparoscopic cholecystectomy. The patients were homogenized by ASA score (ASA I and ASA II) and on population basis. Hemodynamic parameters and 4 time-point pulmonary function tests were monitored in both groups. Peritoneal insufflation resulted in significant increase of systemic arterial pressure (23%), mean arterial pressure (23.8%), systemic vascular resistance (65%), pulmonary vascular resistance (90%), and significant reduction of cardiac output (24%) and cardiac index (51%). Pneumoperitoneum caused transient restriction of pulmonary function by reducing the thoracic and lung compliance. Fall of pH concentration, increase of PaCO2 and ET CO2 without any changes of PaO2, SO2, base excess and bicarbonate ions concentrations were the sequelae of CO2 absorption from peritoneal cavity. Postoperative "hypothermi", i.e. lowering of body temperature for 0.3 degrees C was the consequence of sudden gas expansion (Joule-Thompson phenomenon), which implies continuous flow of dry gas under pressure over peritoneal surface. Tissue damage factors (D-dimer, C-reactive protein, Protein C) were significantly lower in laparoscopic group, meaning that such mode of treatment resulted in minor postoperative pain and shorter period of recovery. Laparoscopy is a revolution in surgery. Definite success of any laparoscopic intervention depends on anesthesia as its crucial factor, at the same time meeting the patient's wish and expectations to be free from pain and discharged in no time from hospital.

Regional hemodynamics after chronic nitric oxide inhibition in spontaneously hypertensive rats.

BACKGROUND: Inhibition of nitric oxide (NO) synthase by L-arginine analogs is associated with elevation of blood pressure in rats. Because endothelium-dependent vasomotion in different vascular beds is not homogenous, the aim of this study was to characterize and compare regional hemodynamic responses in carotid, femoral, and renal vascular beds after chronic NO inhibition in spontaneously hypertensive rats. The possible role of circulating endothelin and renin angiotensin systems in mediating the effects of chronic NO inhibition was also studied. METHODS: Systemic and regional hemodynamics, left ventricular mass, plasma renin activity, and plasma endothelin-1 were determined in control and Nomega-nitro-Larginine methyl ester (L-NAME)-treated (10 mg/kg/day, 4 weeks) spontaneously hypertensive rats. RESULTS: L-NAME treatment increased arterial pressure and total peripheral and regional vascular resistance and decreased cardiac output, stroke volume, and regional blood flow. An increase in blood flow ratio and a decrease in vascular resistance ratio between carotid and renal as well as femoral and renal vascular beds in rats treated with L-NAME was found. Blood flow and vascular resistance ratios between femoral and carotid vascular beds remained unchanged. L-NAME increased plasma renin activity and left ventricular weight/body weight ratio, whereas plasma endothelin-1 was not modified. CONCLUSIONS: The results of this study showed that the renal circulation seemed to be more sensitive to the effects of chronic NO inhibition than carotid and femoral vascular beds. Simultaneous activation of the renin angiotensin system may further potentiate cardiovascular effects of chronic NO inhibition. No evidence that circulating endothelin-1 plays a role in this model of hypertension was found.

Plasma cytokine concentration and the cytokine producing ability of whole blood cell cultures from healthy females with pharmacologically induced hyperprolactinemia.

We investigated the in vitro effect of domperidone-induced hyperprolactinemia on plasma cytokine concentration and blood leukocyte cytokine production in healthy female volunteers. No changes were found in the plasma concentration of interferon (IFN)-gamma, tumor necrosis factor (TNF)-alpha, interleukin (IL)-4, IL-10, IL-6 and IL-13 during hyperprolactinemia when compared with control values. Using unseparated blood leukocytes, we found that the spontaneous production of IL-6 (4-8 h) and transforming growth factor (TGF)-beta 1 (2-4 h) was significantly decreased and that the in vitro stimulated production of IFN-gamma (2-8 h) and TNF (4 h) was significantly increased compared with control. Our data concerning the increased IFN-gamma and TNF producing capacity of unseparated leukocytes during pharmacologically induced hyperprolactinemia strongly support the possibility that the lymphocyte production of these cytokines can be rapidly amplified by prolactin via a priming mechanism.

The acidophilic nature of neuronal Golgi impregnation.

The mechanisms of Golgi impregnation of neurons has remained enigmatic for decades. Recently, it was suggested that divalent (di)chromate anions play a role in the Golgi impregnation process. Therefore, we incubated slices of (para)formaldehyde-fixed rat brain tissue in solutions of potassium (di)chromate, phosphate, chloride or nitrate at pH 6 or 7. Slices were then immersed in solutions of silver nitrate and processed for light microscopical analysis. At pH 6, dichromate probes resulted in dense and homogeneous impregnation of neuronal cytoplasm (typical impregnation). At pH 7, chromate probes showed solely partial cytoplasmic and heavy nuclear-region neuron impregnation (atypical impregnation). Phosphate probes at pH 6 resulted in typical impregnation, whereas at pH 7 phosphate probes gave atypical impregnation. Both at pH 6 and 7, chloride and nitrate probes did not yield any Golgi impregnation. These findings confirmed the pH-dependence of silver-chromate Golgi impregnation as well as the correctness of corresponding acidic silver-phosphate impregnation. Our study revealed a previously unknown, strong anion-dependence of Golgi impregnation, suggesting that hydrogenated monovalent anions are carriers of the neuron impregnation.

[The epithelium of the periodontal pocket--an electron microscopy analysis]. / Epitel parodontalnog dzepa--analiza elektronskim mikroskopom.

The aim of this study was to examine intra- and intercellular changes that characterize the periodontal pocket epithelian. Biopsy specimens of human gingiva were prepared for an electron microscope study by a routine histological procedure. Results of the histological preparations analyses revealed changes significant for pathogenesis of gingivitis and periodontal disease because they cause destruction of epithelial barrier. Such a damaged periodontal pocket epithelium enables further invasion of harmful noxae and spreading of destructive processes into deeper parts of the periodontium.

The effects of long-term low-protein intake on gastrin cells of the rat antral mucosa during adulthood.

The effect of experimental protein malnutrition on gastrin producing cells in the antral part of the stomach was studied in male Wistar rats. Isoenergetic diets containing 25% (C-25) or 6% (PD-6) were given in isocaloric amounts during a 4-month experiment. All rats were offered drinking water ad libitum. The results showed that the long-term protein diet did not produce changes in the gastrin cell number. At the ultrastructural level G cells exhibited a decreased size of the nucleus. They were found to have an increased total granule volume density but the volume density of dense-cored granules was lower. The serum gastrin levels were significantly lowered by feeding the low protein diet. These changes are compatible with decreased functional activity of G cells under long-term protein deprivation.

Rat pancreatic B-cells after chronic alcohol feeding. A morphometric and fine structural study.

Quantitative analysis of the light microscopic and fine structure of rat islet B-cells was carried out in chronic alcoholism. Absolute pancreatic weight and volume were similar in groups C (control) and E (ethanol), but relative pancreatic weight in group E rat was decreased. The results for fasting blood glucose and insulin levels were similar in the two groups of animals. There was a significantly reduced total pancreatic islet volume in E rats. The total number of endocrine cells both per islet and per microns2 of islet was similar in the two groups of animals. The volume density and number of B-cells per islet and per microns2 of islet were not changed in ethanol-treated rats as compared with the control. On the other hand, diameter, surface area and volume of the B-cells and their nuclei were found to be statistically significantly decreased. Histological examination revealed that islet blood vessels were dilated in alcoholic rats. Over the 4-month period of ethanol intake a significant decrease in cell profile area, nuclear profile area and volume density of cytoplasmic granules and an increase in the profile area and volume density of endoplasmic reticulum occurred. The gross histological alteration seen in most B-cells of the ethanol-treated rats was irregularity of the nuclear envelope with deep invagination and with margination of heterochromatin and many empty granules or granules without clear electron dense crystals of insulin. The present results indicate some optical and structural abnormalities of B-cells in chronic alcoholism that may be related to cell dysfunction and may contribute, at least in part, to the endocrine pancreas functional disturbance.

Effect of domperidone-induced hyperprolactinemia on selected immune parameters in healthy women.

Domperidone, anti-emetic drug, given to healthy female volunteers, induced an elevation of plasma prolactin (PRL) concentration with the peak in 1-4 h. The release of prolactin had a transient stimulating effect on theophylline sensitive T lymphocytes and on concanavalin A induced mitogenic activity, suggesting an enhanced activity of T suppressor lymphocytes. The relative number of CD4+ lymphocytes decreased markedly one hour after domperidone administration and returned to normal values within 2 h (that means 3 h after taking the drug). The number of lymphocytes positive for dipeptidyl peptidase IV exhibited similar transient increase and normalization of activity. No change was observed in the number of CD8+ lymphocytes. The production of interferon by leukocytes treated with Newcastle disease virus was found to be significantly increased 2 h after domperidone administration. The results suggest that prolactin can selectively stimulate some functions of cellular immunity as well as the release of cytokines (IFN). The present study may contribute to the understanding of the role of the immune system in endogenous hyperprolactinemia.

Effect of chronic alcohol feeding on the ultrastructure of rat peripheral blood neutrophils: a morphometric study.

Morphometric methods were used to analyze the ultrastructural characteristics of peripheral blood polymorphonuclear neutrophils (PMN) in 10 rats chronically consuming ethanol and 20 rats fed an isoenergetic standard diet (10 ad libitum and 10 pair fed control rats). Morphometric measurements were made, after a 4-month experimental period, of the following: the profile area of the cell, nucleus and cytoplasm; nucleus to cell profile area; volume density of the nucleus, cytoplasm, mitochondria, Golgi system, endoplasmic reticulum and cytoplasmic granules; number of mitochondria per cell profile; number of cytoplasmic granules per cell profile and per micron2 of cytoplasm, as well as the azurophilic to specific granule ratio and mean diameter of granules. A significant decrease in cell profile area and cytoplasm profile area was shown in ethanol-treated rats. The volume density of mitochondria and endoplasmic reticulum nearly doubled during ethanol abuse. The results also showed that there were highly significant effects of ethanol on the total number of cytoplasmic granules per cell. In addition, changes were observed in mitochondria such as clumping, elongation, swelling and disruption of cristae, as well as changes in the topographic distribution of granules in the cytoplasm such as registration of cytoplasmic areas with numerous granules and areas with a smaller number or without any granules. Some neutrophils of ethanol-treated rats had autophagic vacuoles. The results indicate some ultrastructural abnormalities of PMN in chronic experimental alcoholism that may be related to polymorphonuclear phagocyte dysfunction.

[New findings on the immunologic diagnosis of systemic lupus erythematosus]. / Nové poznatky v imunologickej diagnostike systémového lupus erythematosus.

The authors review recent findings pertaining to the pathogenesis of systemic lupus erythematosus. It was revealed that knowledge of impaired humoral and cellular immunity is of great practical importance and substantially improves the prognosis of the disease.

Effects of chronic ethanol administration on the serotonin-producing cells in rat gastric antral and duodenal mucosa.

The present study describes our observations on optical and ultrastructural features of serotonin-containing cells in the rat antral and upper duodenal mucosa, utilizing optic morphometric measurements in a model of experimental chronic alcoholism of rat in which nutrition was well controlled. Male Wistar rats were given ethanol to provide 23 per cent of the total calories, while starch replaced ethanol isocalorically in controls. Twenty-five per cent of the calories were provided by protein in both groups. Blood levels of serotonin were significantly raised after chronic ethanol feeding (0.059 +/- 0.06 vs. 0.159 +/- 0.012 micrograms/ml, p < 0.01). Decrease in the number of immunohistochemically-detectable serotonin-containing cells was found in the pyloric gland mucosal area specimens of the chronically ethanol-treated rats (68.9 +/- 5.2 vs 43.3 +/- 3.0; p < 0.001). The immunohistologically-evaluated number of the same cells in the duodenal mucosa specimens was significantly decreased by alcohol feeding. Although total villi and crypt count per whole circular section, and the number of crypts per villus were not significantly changed either in control animals or in chronically ethanol-fed rats, decreased number of these cells per whole circular section (289 +/- 21.6 vs. 183 +/- 10.5; p < 0.001) per villus (2.52 +/- 0.14 vs. 1.21 +/- 0.10; p < 0.001) and per crypts (0.97 +/- 0.08 vs. 0.79 +/- 0.04; p < 0.05) were reported after alcohol consumption. In both control and experimental rats the cells were predominantly found in the basal half of the antropyloric mucosa. Alcohol did not lead to any changes in normal distribution of the duodenal serotonin-producing cells. The above quantitative changes in serotonin-producing cells were not accompanied by changes in their subcellular appearance in stomach and duodenal mucosa of alcohol-treated rat.

[Use of the modified Warthin-Starry method in pigmented nevi of the skin]. / Primena modifikovane Warthin-Starryjeve metode kod pigmentnih nevusa koze.

Warthin-Starry method is an argyrophilic method, performed at ph 3.2. It is a very efficient method for demonstration of melanin pigment granules in the nevus tissue and in malignant melanoma. We have concluded that it is highly specific for demonstration of melanin pigment granules, and more efficient than the Fontan-Masson method. The Fontan-Mason method reduces other pigments in the cell, apart from melanin. Modified Warthin-Starry method at ph 3.2 in the nevus tissue reduces only melanin pigment granules.

[Ultrastructure of pigmented skin nevi]. / Ultrastruktura pigmentnih nevusa koze.

Nevus cells are biologically significant because they are associated with epidermal melanocytes, while their clinical malignant melanomas. Apart from many other ultrastructural features their mutual characteristic is that they are exceptionally diverged. The main feature distinguishing nevus cells from melanocytes is the presence of big binuclear cells as well as frequent occurrence of grouped melanosomes in their cytoplasm.

A comparison of regulatory cells in spinal fluid and blood in patients with multiple sclerosis and other neurologic diseases.

Multiple sclerosis is a disease in which immune abnormalities are present both in the CNS and peripheral blood. Whether these changes are primary or secondary to the disease process is not known. We tested T-cell clones derived from activated lymphocytes in the blood and CSF of MS patients and controls for their capacity to regulate T-cell responses to alloantigens. A wide spectrum of regulatory functions were observed, ranging from marked enhancement to almost complete suppression. Clones from different patient populations and anatomic sites were equivalent in their regulatory functions with the net effect of clones in each compartment being suppression. However, certain clones from CSF and peripheral blood had the capacity to stimulate autologous T cells. Percentages of such clones in the peripheral blood of MS patients were significantly higher than in controls, while percentages in MS and other neurologic diseases (OND) CSF were equivalent. Our data suggest that (1) functional suppressor cells are not lost from the blood or CSF or MS and OND patients, (2) lymphocytes that have entered the CNS in patients with MS and other CNS diseases have equivalent regulatory functions, (3) MS may be an illness in which peripheral immunologic events are important in perpetuating the disease process, and (4) responses to autologous antigens may also play a role in this perpetuation.

[Immunobiologic properties of lactobacilli]. / Imunobiologické vlastnosti laktobacilov.

The authors investigated in experiments on mice immunobiological properties of selected strains of lactobacilli (Lbc. acidophilus, Lbc. casei and Lbc. delbruecki). Their immunostimulating action was evaluated from the migrating capacity of lymphocytes into the interepithelial spaces and lamina propria mucosae of the gut. The most marked changes were observed in the group of animals to whom Lbc. acidophilus and Lbc. casei was administered for two weeks by a gastric tube. The protective properties of lactobacilli on the course and development of model infections (virus of encephalomyocarditis) was greatest in mice given Lbc. casei and Lbc. acidophilus by the intraperitoneal route four days before infection. At the end of the two-week period in the Lbc. casei group 66% mice survived and in the Lbc. acidophilus group 34%. The ability of lactobacilli to influence the interferon producing activity was investigated in vitro on a model of peritoneal cells obtained from premedicated mice. The lactobacilli strains themselves did not have interferon inducing properties. However, when the interferon producing capacity of peritoneal cells was assessed after administration of the viral inducer (virus of Newcastle disease) the capacity was much higher, when compared with controls.

Human acid- and thermolabile alpha-interferon-like substance: selective reactivity with a monoclonal antibody.

An acid- and thermolabile alpha-interferon-like substance, designated AL-IFN-alpha, has been found in non-processed normal human leukocyte IFN preparations as well as sera from patients with autoimmune or other chronic diseases. Little is known about origin, production and biological activity of these IFN activities. Monoclonal antibodies were obtained which proved highly selective in neutralizing AL-IFN-alpha in both anti-proliferative and antiviral tests. While the monoclonal antibodies were strict specific, polyclonal antibodies against various interferons showed less specificity in these tests. The results suggest that AL-IFN-alpha represents an antigenically distinct IFN-alpha subtype or, alternatively, a new lymphokine with antiproliferative and antiviral activity.

Abnormal macrophages and NK cell cytotoxicity in human systemic lupus erythematosus and the role of interferon and serum factors.

Macrophage (MO) and natural killer (NK) cell mediated cytotoxicity to K562 target cells were strikingly decreased in patients with systemic lupus erythematosus (SLE). SLE NK cells failed to release soluble factor(s) for lysing the targets. IFN-induced enhancement of both types of cytotoxicity was impaired. NK cells from healthy subjects kept their activity in culture with or without IFN for more than six days whereas SLE NK cell activity declined to zero at day 3. So, the increased IFN level of many SLE patients and a possible prior IFN priming effect seemed unrelated to the insensitivity to exogenous IFN in vitro. Inhibition factor(s) of SLE serum suppressed NK cytotoxicity in the presence of IFN whereas IFN sensitivity of MO remained unaffected indicating the complex regulation by serum components of immune reactions.