Giant conjunctival melanoma treated with excision and primary reconstruction using a partial thickness skin graft.

Conjunctival melanoma is a rare type of malignancy of ocular surface tissue with a high recurrence rate, sight, and life-threatening malignancy. In this paper, we report a case of giant conjunctival melanoma in an 86-year-old woman, localized on her left eye. The pigmented tumor lesion was de novo occurred and treated by local wide excision and reconstruction of the presented defect by partial thickness skin from the inside parts of the forearm. The eye itself was not damaged, and the first-line lymph nodes showed no malignant activity. In the three years of follow-up, no signs of recurrence have been observed. The treatment approach with local excision and subsequent reconstruction with the partial thickness skin graft or excision with adjuvant therapy has given a good clinical outcome.

Maternal mortality in Serbia.

Maternal mortality related to obstetric events is still high today. The main components of reproductive health are evaluated. Early diagnosis of the obstetric risks can significantly reduce maternal mortality.

KLF4 expression and apoptosis-related markers in gastric cancer.

PURPOSE: To correlate the expression of Kruppel-like factor 4 (KLF4) with clinicopathological properties of gastric cancer (GC) and to evaluate any possible correlation between KLF4 expression and the expression of apoptosis-related markers p53, Fas, Bcl-2, survivin and FLICE inhibitory protein (Flip-l). METHODS: Formalin-fixed, paraffin-embedded tissue specimens obtained from 96 patients with GC who had undergone gastric surgery were analyzed for pathological parameters, while KLF4, p53, Fas, Bcl-2, survivin and Flip-l expression was assessed by immunohistochemistry. RESULTS: TKLF4 immunohistochemical staining was noted in 78.1% of the cases. Strong positivity was found in 15.6% and weak in 62.5% of the samples. Positive expression of p53, Fas, Bcl-2, survivin, Flip-l was found in 56.2%, 44.8%, 15.6%, 41.7% and 38.5% of the samples, respectively. KLF4 expression was significantly associated with p53 nuclear staining and Fas immunoreactivity. p53-positive tumors demonstrated more often high KLF4 staining compared to p53-negative tumors. Fas-positive tumors were associated with decreased KLF4 expression. Logistic regression analysis of apoptosis-related markers to KLF4 expression revealed that Fas positivity significantly decreased the probability of strong KLF4 expression, and inversely, Bcl-2 expression improved the prediction of KLF4 staining. When all 5 predictive variables were considered together (p53, Fas, survivin, Bcl-2, Flip-l) they significantly predicted the type of KLF4 expression in GC cells (p=0.019). CONCLUSION: Our results suggest that the decrease or loss of KLF4 expression correlates with diffuse-type GC and immunoreactivity to Fas, and are inversely linked with p53 nuclear accumulation. The significance of KLF4 in GC requires further studies and should be more thoroughly investigated for potential use in the evaluation and better stratification of GC patients.

Endoscopic ultrasound for differential diagnosis of duodenal lesions.

PURPOSE: Duodenal tumors are rare and require a different management from that of esophagogastric neoplasia. The present study retrospectively analyses the endoscopic ultrasound (EUS) features of duodenal tumors of both epithelial and subepithelial origin. MATERIALS AND METHODS: During a 12 year period, all duodenal tumors with histologic confirmation by surgery or biopsy were collected including endoscopic and endosonographic images. EUS images were analyzed for specific features (echogenicity, wall layer structure and relation, outer margins) to possibly distinguish epithelial (polyps and carcinoma versus lymphoma) and subepithelial (tumor type) tumors. RESULTS: 53/80 cases had histologic confirmation (mean age 53.1 ±â€Š11.4 years, m:f = 33:20), 31 were epithelial (13 adenomas, 12 carcinomas, 6 lymphomas) and 22 subepithelial (11 GISTs, 7 Brunneromas, 1 lipoma, 3 NETs). EUS did not recognize carcinomas in 2/13 adenomas. EUS features suggesting carcinoma were loss of wall layers and irregular margins. 5/6 lymphomas showed inhomogeneous thickening with layers partially recognizable. Tumor type of subepithelial lesions correlated with echogenicity: GIST tumors were mostly (62.5 %) hypocheoic with the 3 malignant cases being characterized by heterogeneous echopattern with irregular outer margins. Of the hyperechoic lesions, lipomas had a homogeneous whitish appearance, while NET and Brunneromas were less hyperechoic. In the latter, the endoscopic aspect was also helpful for differential diagnosis. Accuracy of combined endoscopic/EUS imaging for all duodenal lesions was 84.9 % (45/53). No procedural complications occurred among all patients that received EUS examinations. CONCLUSION: EUS contributes to the differential diagnosis of epithelial lesions known to be malignant; in subepithelial tumors, tissue confirmation is still required.

Gastrointestinal and liver tumours and public health in Europe.

STATE OF THE ART: Europe comprises only one eighth of the total world population but has around one quarter of the global total of cancer cases--some 3.2 million new patients per year. While the disproportionate cancer burden is readily apparent, the disease patterns in Europe cannot simply be generalized--overall cancer incidence and mortality rates vary at least two-fold between European countries and the differences are often far greater for specific cancers. With 1.7 million deaths each year, cancer currently represents the second most important cause of death in Europe. The range of survival rates is similarly wide. For individual cancers, the variation across Europe is even greater. This reflects a wide range of social and epidemiological factors in different countries: cancer prevention programmes; screening programmes; cancer control plans; individual lifestyles and occupational exposures; the existence and accessibility of health-care facilities and technological infrastructure; and the availability of human, financial and material resources for health and economic development. Europe has some of the richest countries in the world, but also some of the poorest. In 2002, 168 million people were living below the poverty line, about 46% of the European population. The time trends in cancer risk also vary between European countries and some cancers show different trends between men and women, or young and old, or poor and rich. The public health profile of cancer in Europe is complex. Trends in the incidence and mortality rates are also influenced by successes in health promotion (e.g. tobacco control), efficient screening (e.g. breast, bowel, cervix) and better treatment. These have been reflected in lower incidence, reduced mortality, higher survival, improved life expectancy and a better quality of life for cancer survivors. CONCLUSIONS: Cancer of the gastrointestinal (GI) tract is the most common cancer in Europe. More than half of GI cancer cases arise from the colon. They can remain asymptomatic until late in the natural history of the disease, and as this is the stage at which they can be cured, screening has been advocated for well members of the population and surveillance for those with conditions predisposing to cancer.

The Regenerative Potential of Donkey and Human Milk on the Redox-Sensitive and Proliferative Signaling Pathways of Skin Fibroblasts.

The influence of milk bioactive peptides on skin regenerative potential and rejuvenation is very often limited because of allergic reactions. The current study is aimed at exploring the influence of donkey colostrum and mature milk, human colostrum and mature milk, and ß-casein and ß-casomorphine-7, on the growth and inflammatory response of the culture of cultured skin fibroblasts exposed to these conditions for twenty-four hours. Their effects on the growth-regulatory kinases and redox-sensitive, proinflammatory transcriptional factor NF-κB were detected by using specific primary antibodies against NF-κB p65, Akt-1, phospho-Akt-1, Erk-1, phospho-Erk-1, JNK, phospho-JNK, phospho-STAT-1, and CD26, while logarithmic integrated fluorescence intensity patterns were recorded by flow cytometry. The downregulation of NF-κB p65 was observed after the exposure of skin fibroblasts to donkey milk and human colostrum, while ß-casein and ß-casomorphine-7 exerted the opposite effect, which suggests that noncasein bioactive peptides of donkey and human milk may be responsible for anti-inflammatory properties. The exposure to all milk species examined and ß-casein leads to the activation of growth-regulatory kinases (Akt1/2/3 kinase, Erk kinase, JNK kinase, and Stat-1 kinase), especially for the p-Erk pathway, which suggests that essential amino acids of casein may be responsible for Erk-induced cell cycle activation and proliferation. The opposite effect was observed when cells were exposed to ß-casomorphine-7, which may affect the skin fibroblast survival and their proliferative and regenerative potential. Donkey milk did not significantly change the CD26 antigen expression. In conclusion, our results suggest that among cell signaling molecules, the most sensitive but nonspecific downstream effector is p-Erk kinase, which may point to donkey milk usefulness in wound healing, regenerative, and aesthetic dermatology. The noncasein bioactive peptides of donkey milk may be responsible for the anti-inflammatory property of donkey milk and colostrum, which may indicate the usefulness in the treatment of inflammatory skin diseases.

Feeding produced in the satiated rat by elevating the concentration of calcium in the brain.

When the concentration of calcium ions in the cerebral ventricles is elevated, a fully satiated rat eats voraciously. This feeding response is not prevented by prior intraventricular administration of alpha-or beta-adrenergic blocking agents, or other pharmacological antagonists. This supports the concept of an independent ionic mechanism, rather than a neurotransmitter one, for modulating a "set-point" for weight or hunger.

Triple-way therapeutic approach for paraganglioma-dependent erythrocytosis: drugs and surgery plus "multi-manner" apheresis.

Paragangliomas are tumors arising from the extra-adrenal paragangliar neural crest cells. The sympathoadrenal neuroendocrine system consists of extra-adrenal paragangliar cellular layer along the paravertebral and para-aortic axis, and the adrenal medullae. Paraganglioma should be included in the differential diagnosis of secondary erythrocytosis due to its possible ectopic erythropoietin (EPO) secretion. Thus, in this report we present a 24-year-old female patient with onset of unregulated ectopic EPO secretion, and consecutive erythrocytosis followed by hypertension, secondary to paraganglioma of multifocal retroperitoneal localization. Clinical, laboratory, and radiological investigations confirmed both an elevated EPO level and the presence of multiple paraganglioma. This paraneoplastic-mediated medical condition with high risk of cellular hyperviscosity syndrome (CHVS) requires prompt diagnosis and rapid therapeutic interventions. Initially, simple phlebotomy procedures were used; following that, tumors were surgically removed. In the course of the disease, this tumor relapsed, and urgent apheresis, as a treatment of life-threatening state, was used. The therapy performed resulted in a rapid blood viscosity depletion and a significant (P < 0.01) serum EPO reduction, as well as the general clinical benefit. Therefore, we conclude that the use of our own "multi-manner" apheresis (erythrocythapheresis plus plasma exchange), for long-time interval (until further causative therapy), effectively cross-bridged the possible hazards of EPO-dependent CHVS.

Mitogen-activated and cyclin-dependent protein kinases selectively and differentially modulate transcriptional enhancement by the glucocorticoid receptor.

Cyclin-dependent kinase (CDK) and mitogen-activated protein kinase (MAPK) phosphorylate the rat glucocorticoid receptor in vitro at distinct sites that together correspond to the major phosphorylated receptor residues observed in vivo; MAPK phosphorylates receptor residues threonine 171 and serine 246, whereas multiple CDK complexes modify serines 224 and 232. Mutations in these kinases have opposite effects on receptor transcriptional activity in vivo. Receptor-dependent transcriptional enhancement is reduced in yeast strains deficient in the catalytic (p34CDC28) or certain regulatory (cyclin) subunits of CDK complexes and is increased in a strain devoid of the mammalian MAPK homologs FUS3 and KSS1. These findings indicate that the glucocorticoid receptor is a target for multiple kinases in vivo, which either positively or negatively regulate receptor transcriptional enhancement. The control of receptor transcriptional activity via phosphorylation provides an increased array of regulatory inputs that, in addition to steroid hormones, can influence receptor function.

Peptidomics of an in vitro digested α-Gal carrying protein revealed IgE-reactive peptides.

The mammalian carbohydrate galactose-α1,3-galactose (α-Gal) causes a novel form of food allergy, red meat allergy, where patients experience severe allergic reactions several hours after red meat consumption. Here we explored gastric digestion of α-Gal glycoproteins using an in vitro model. Bovine thyroglobulin (BTG), a typical α-Gal carrying glycoprotein, was digested with pepsin. The resulting peptides were characterised by SDS PAGE, immunoblot and ImmunoCAP using sera from 20 red meat allergic patients. During pepsinolysis of BTG, a wide range of peptide bands was observed of which 14 to 17 kDa peptides remained stable throughout the gastric phase. The presence of the α-Gal epitope on the obtained peptides was demonstrated by an anti-α-Gal antibody and IgE from red meat allergic patients. The α-Gal digests were able to inhibit up to 86% of IgE reactivity to BTG. Importantly, basophil activation test demonstrated that the allergenic activity of BTG was retained after digestion in all four tested patients. Mass spectrometry-based peptidomics revealed that these peptides represent mostly internal and C-terminal parts of the protein, where the most potent IgE-binding α-Gal residues were identified at Asn1756, Asn1850 and Asn2231. Thus allergenic α-Gal epitopes are stable to pepsinolysis, reinforcing their role as clinically relevant food allergens.

Peanut protein structure, polyphenol content and immune response to peanut proteins in vivo are modulated by laccase.

Food texture can be improved by enzyme-mediated covalent cross-linking of different food components, such as proteins and carbohydrates. Cross-linking changes the biological and immunological properties of proteins and may change the sensitizing potential of food allergens. In this study we applied a microbial polyphenol oxidase, laccase, to cross-link peanut proteins. The size and morphology of the obtained cross-linked proteins were analyzed by electrophoresis and electron microscopy. Structural changes in proteins were analyzed by CD spectroscopy and by using specific antibodies to major peanut allergens. The bioavailability of peanut proteins was analyzed using a Caco-2 epithelial cell model. The in vivo sensitizing potential of laccase-treated peanut proteins was analyzed using a mouse model of food allergy. Finally, peanut polyphenols were analyzed by UHPLC-MS/MS, before and after the enzymatic reaction with laccase. Laccase treatment of peanut proteins yielded a covalently cross-linked material, with the modified tertiary structure of peanut proteins, improved bioavailability of Ara h 2 (by 70 fold, p < 0.05) and modulated allergic immune response in vivo. The modulation of the immune response was related to the increased production of IgG2a antibodies 11 fold (p < 0.05) and reduced IL-13 secretion in in vitro cultured splenocytes 7 fold (p < 0.05). Analysis of the peanut polyphenol content and profile by HPLC-MS/MS revealed that laccase treatment depleted the peanut extract of polyphenol compounds leaving mostly isorhamnetin derivatives and procyanidin dimer B-type in detectable amounts. Treatment of complex food extracts rich in polyphenols with laccase results in both protein cross-linking and modification of polyphenol compounds. These extensively cross-linked proteins have unchanged potency to induce allergic sensitization in vivo, but certain immunomodulatory changes were observed.

Pressor response mediated via histamine H1-receptors of the guinea-pig sympathetic ganglia.

In pithed guinea-pigs, the general characteristics and origin of the pressor response to intravenous injection of histamine were examined. Histamine (5-80 micrograms/kg) produced a rapid, short-lasting, constant, prominent and dose-dependent pressor response, followed by a secondary slight and prolonged depressor response. The vascular response to histamine was accompanied by a marked tachycardia. The pressor effect of histamine (30 micrograms/kg) was strongly reduced or abolished in animals pretreated with nicotine, reserpine, bretylium or 6-hydroxydopamine. Furthermore, pyrilamine, a histamine H1-receptor antagonist, antagonized in a dose-dependent manner the pressor response to histamine. On the contrary, metiamide, a histamine H2-receptor antagonist, as well as hexamethonium and atropine, cholinergic antagonists, did not suppress the pressor effect of histamine. The present experiments provide evidence that in guinea-pigs, the pressor component of the vascular response to histamine results predominantly from the activation of histamine H1-receptors in the sympathetic ganglia with consequent release of noradrenaline at postganglionic sympathetic nerve terminals.

A further study of the cardiovascular responses to central administration of acetylcholine in rats.

General characteristics and the nature of cardiovascular responses to acetylcholine (ACh) injected into the lateral cerebral ventricle (LCV), the 4th cerebral ventricle (4th CV) and the cisterna magna (CM) were studied. Acetylcholine, depending on the route of injection, the dose and the peripheral vascular tone, produced both increases and decreases in blood pressure and heart rate. After injection of central anticholinesterase agents (physostigmine, neostigmine, armin), the pressor response to ACh was reversed to a depressor response, with or without a secondary pressor response, accompanied by a bradycardia. The vascular effects were selectively abolished by central administration of atropine, indicating that they are mediated via muscarinic cholinoceptors. It is concluded that cardiovascular responses to ACh can be initiated at a number of cardiovascular loci situated at various levels of the rat brain. It seems that both muscarinic cholinoceptors mediating pressor responses and those mediating depressor responses exist in the cardiovascular loci of all these brain levels, but not in an equal proportion. The hypothesis is proposed that the muscarinic cholinoceptors responsible for pressor effects are more sensitive to ACh and functionally probably more competent than muscarinic cholinoceptors responsible for depressor effects, the latter being in a markedly greater number. The present study provides additional support for the idea that ACh participates in the physiological control of the rat cardiovascular system, as both a central cholinergic excitatory and inhibitory regulator.

Analysis of the cardiovascular responses to central injection of tryptamine in rats.

Tryptamine (2-20 micrograms), administered into the lateral cerebral ventricle of the rat, evoked a pressor response which was sometimes followed by a prolonged depressor response. The intracisternal administration of tryptamine (7-20 micrograms) caused a slow progressive and long-lasting depressor effect without or with an initial pressor effect. The pressor response was accompanied by variable changes in heart rate, whilst the pure depressor response was accompanied by a decrease in heart rate. After transection of the spinal cord between C1 and C2 the pressor response was substantially reduced or abolished. Methysergide, injected centrally, antagonized in a dose-dependent manner the pressor effect, whilst p-chlorophenylalanine, atropine and hexamethonium, administered by the same route, did not diminish this effect. It is concluded that tryptamine, injected centrally, causes both increases and decreases in arterial blood pressure and heart rate. The pressor response to tryptamine results from the activation of central noncholinergic, methysergide-sensitive, receptor sites and the depressor response to tryptamine may be due to a centrally-induced reduction in sympathetic nervous activity. It is tentatively suggested that tryptamine, like 5-hydroxytryptamine, participates in the physiological regulation of the cardiovascular system of the rat, as both a central excitatory and inhibitory regulator.

Divergent effects of cyproheptadine and R 50 656, a 5-HT2 antagonist, on the cardiovascular response to 5-hydroxytryptamine in rats.

Cyproheptadine and R 50 656, a novel 5-HT2 antagonist, produced a dose-dependent suppression of the pressor response to 5-hydroxytryptamine (5-HT), but did not affect the depressor response. The pressor effect of 5-HT was significantly more sensitive to R 50 656 than to cyproheptadine. On the contrary, the tachycardic response to 5-HT was antagonized only with cyproheptadine. Accordingly, in rats the 5-HT receptors of cardiac tissue differ in their pharmacological properties from both the excitatory and inhibitory 5-HT receptors of vascular smooth muscle.

Characterization of visually similar diffuse diseases from B-scan liver images using nonseparable wavelet transform.

This paper describes a new approach for texture characterization, based on nonseparable wavelet decomposition, and its application for the discrimination of visually similar diffuse diseases of liver. The proposed feature-extraction algorithm applies nonseparable quincunx wavelet transform and uses energies of the transformed regions to characterize textures. Classification experiments on a set of three different tissue types show that the scale/frequency approach, particularly one based on the nonseparable wavelet transform, could be a reliable method for a texture characterization and analysis of B-scan liver images. Comparison between the quincunx and the traditional wavelet decomposition suggests that the quincunx transform is more appropriate for characterization of noisy data, and practical applications, requiring description with lower rotational sensitivity.

Graphic assessment of interincisal point movements during chewing of hard and soft foods.

Interincisal point movements in 35 healthy, dentate subjects were investigated with an electrognathic apparatus. The main focus of the study was to find the average values of determined point graphic assessment in frontal and sagittal planes during chewing of hard and soft foods. However, as this was insufficient to obtain information on areas where occlusal contacts were reached, border movement envelopes were also recorded to obtain indirect information in connection to the previous task. The results showed that the graphic assessment form of mandibular movements was dependent on the consistency of the food that was chewed.