Rilotumumab plus epirubicin, cisplatin, and capecitabine as first-line therapy in advanced MET-positive gastric or gastro-oesophageal junction cancer (RILOMET-1): a randomised, double-blind, placebo-controlled, phase 3 trial.

BACKGROUND: Rilotumumab is a fully human monoclonal antibody that selectively targets the ligand of the MET receptor, hepatocyte growth factor (HGF). We aimed to assess the efficacy, safety, and pharmacokinetics of rilotumumab combined with epirubicin, cisplatin, and capecitabine, and to assess potential biomarkers, in patients with advanced MET-positive gastric or gastro-oesophageal junction adenocarcinoma. METHODS: This multicentre, randomised, double-blind, placebo-controlled, phase 3 study was done at 152 centres in 27 countries. We recruited adults (aged ≥18 years) with unresectable locally advanced or metastatic gastric or gastro-oesophageal junction adenocarcinoma, an Eastern Cooperative Oncology Group (ECOG) performance status of 0 or 1, MET-positive tumours (≥25% of tumour cells with membrane staining of ≥1+ staining intensity), and evaluable disease, who had not received previous systemic therapy. Eligible patients were randomly assigned (1:1) via a computerised voice response system to receive rilotumumab 15 mg/kg intravenously or placebo in combination with open-label chemotherapy (epirubicin 50 mg/m2 intravenously; cisplatin 60 mg/m2 intravenously; capecitabine 625 mg/m2 orally twice daily) in 21-day cycles for up to ten cycles. After completion of chemotherapy, patients continued to receive rilotumumab or placebo monotherapy until disease progression, intolerability, withdrawal of consent, or study termination. Randomisation was stratified by disease extent and ECOG performance status. Both patients and physicians were masked to study treatment assignment. The primary endpoint was overall survival, analysed by intention to treat. We report the final analysis. This study is registered with ClinicalTrials.gov, number NCT01697072. FINDINGS: Between Nov 7, 2012, and Nov 21, 2014, 609 patients were randomly assigned to rilotumumab plus epirubicin, cisplatin, and capecitabine (rilotumumab group; n=304) or placebo plus epirubicin, cisplatin, and capecitabine (placebo group; n=305). Study treatment was stopped early after an independent data monitoring committee found a higher number of deaths in the rilotumumab group than in the placebo group; all patients in the rilotumumab group subsequently discontinued all study treatment. Median follow-up was 7·7 months (IQR 3·6-12·0) for patients in the rilotumumab group and 9·4 months (5·3-13·1) for patients in the placebo group. Median overall survival was 8·8 months (95% CI 7·7-10·2) in the rilotumumab group compared with 10·7 months (9·6-12·4) in the placebo group (stratified hazard ratio 1·34, 95% CI 1·10-1·63; p=0·003). The most common grade 3 or worse adverse events in the rilotumumab and placebo groups were neutropenia (86 [29%] of 298 patients vs 97 [32%] of 299 patients), anaemia (37 [12%] vs 43 [14%]), and fatigue (30 [10%] vs 35 [12%]). The frequency of serious adverse events was similar in the rilotumumab and placebo groups (142 [48%] vs 149 [50%]). More deaths due to adverse events occurred in the rilotumumab group than the placebo group (42 [14%] vs 31 [10%]). In the rilotumumab group, 33 (11%) of 298 patients had fatal adverse events due to disease progression, and nine (3%) had fatal events not due to disease progression. In the placebo group, 23 (8%) of 299 patients had fatal adverse events due to disease progression, and eight (3%) had fatal events not due to disease progression. INTERPRETATION: Ligand-blocking inhibition of the MET pathway with rilotumumab is not effective in improving clinical outcomes in patients with MET-positive gastric or gastro-oesophageal adenocarcinoma. FUNDING: Amgen.

Evaluation of a hydrophobic gel adhering to the gingiva in comparison with a standard water-soluble 1% chlorhexidine gel after scaling and root planing in patients with moderate chronic periodontitis. A randomized clinical trial.

OBJECTIVES: To evaluate the clinical, microbiological and enzymatic activity of a hydrophobic chlorhexidine-based gingiva-adhering gel containing herbal ingredients, compared with a commercially available 1% chlorhexidine water-soluble gel, during non-surgical therapy of moderate chronic periodontitis. METHODS: A total of 34 subjects participated in this 6-month blinded randomized parallel controlled trial (ISRCTN35210084). After scaling and root planing (SRP), test group received the gel, by rubbing on the gingiva, once every second day, for 14 days. The control group received the control gel twice daily. Clinical parameters considered were the approximal plaque index, simplified oral hygiene index, modified gingival index, bleeding on probing, probing depth and clinical attachment level (primary outcome), assessed at baseline, 3 and 6 months, together with the frequency of detection of Aggregatibacter actinomycetemcomitans, Porphyromonas gingivalis (P.g.), Prevotella intermedia, Treponema denticola (T.d.), Tannerella forsythia (T.f.), and activity of neutrophil elastase and myeloperoxidase (secondary outcomes). RESULTS: At 3 and 6 months, all clinical parameters improved significantly, without significant intergroup differences, except OHI-S, which improved at 3 months (P < 0.05). Microbiological data resulted in no significant intergroup differences at baseline and 6 months. At 3 months, significant differences for P.g., T.f. and T.d. were noted. A significant reduction of neutrophil elastase after 3 and 6 months was observed (P < 0.005), without significant intergroup differences. For myeloperoxidase, significant reductions were noted in both groups (P < 0.005 and P < 0.05), but no significant intergroup differences. The tested product seemed to have an increased efficacy, due to longer persistence on the gingiva, with reduced application frequency. CONCLUSIONS: Both products had a relatively similar influence on the clinical, microbiological and enzymatic outcomes at 3 and 6 months after SRP.

Altered levels of plasma chemokines in breast cancer and their association with clinical and pathological characteristics.

Chemokines are a family of small, structurally related cytokines with chemoattractant and activation properties. In breast cancer, both epithelial cancer cells and cells within the microenvironment secrete chemokines with either tumor-promoting or anti-malignant potential. The equilibrium between these two chemokine activities plays a key role in the biology of the developing tumor, including its ability to metastasize. Here we evaluated the expression of chemokines in breast tumors and the plasma of breast cancer patients before treatment in order to identify a blood-based signature that could distinguish between malignant and non-malignant processes. We screened the mRNA expression of chemokine genes using cDNA microarray on homogenous, laser-capture microdissected breast cancer specimens. Further, using a protein array approach, we determined the levels of selected chemokines in the plasma of patients with breast cancer, benign breast tumors and healthy women. Finally, we analyzed the association between the levels of chemokines in breast and blood samples with the pathological characteristics of the disease. At mRNA level, 27 chemokines and 11 chemokine receptors were differentially expressed in cancers when compared with normal breast tissue. When compared to benign tumors, the only chemokine significantly upregulated in cancers was CXCL10. At protein level, with the exception of CXCL13, nine out of the ten selected chemokines (CCL2, CCL7, CCL18, CCL22, CXCL8, CXCL9, CXCL10, CXCL11 and osteoprotegerin) were significantly overexpressed in the plasma of breast cancers patients compared to healthy controls. After grouping, CXCL8, CXCL9 and CCL22 proved to be significant predictors for breast cancers as compared to healthy controls in a model of logistic regression. We found upregulation of CXCL8, CXCL11 and CXCL9 in triple negative carcinomas, CXCL9 in low proliferative carcinomas, and CXCL10, CCL7 and osteoprotegerin in poorly differentiated carcinomas. Furthermore, CXCL9 was overexpressed in lymph node negative tumors, whereas CXCL8 and CCL18 were higher in advanced stage carcinomas. We identified a panel of chemokines dysregulated in breast cancer that could be further investigated as prospective novel diagnostic markers or for therapeutic and prognostic applications.

Prophylactic Percutaneous Endoscopic Gastrostomy (PEG) - The Importance of Nutritonal Support in Patients with Head and Neck Cancers (HNCs) or Neurogenic Dysphagia (ND).

BACKGROUND: We evaluated the effectiveness and safety of prophylactic PEG performed for the enteral nutrition support during the oncological treatment of patients with HNCs and as a part of the management of neurological patients experiencing neurogenic dysphagia. METHODS: In 2013 we followed up on a group of 23 HNC patients subjected to prophylactic PEG. We assessed the duration of the procedure, intraprocedural incidents and their causes, time to tube-refeeding and discharge after intervention, post interventional analgesia, early and late complications,toleration, costs and postoperative course of these patients after radical surgery maintaining PEG in place. In parallel we followed up on a group of 10 neurological patients who have undergone a PEG placement to improve the nutrional status and to prevent recurrent chest infections due to ND related silent aspiration. RESULTS: The procedures were performed under sedation with Midazolam and the mean duration was about 7 minutes.Postoperative analgesia was minimal. Refeeding through the tube was initiated 2-4h hours later and the patients were discharged 12-24h after the procedure. Early complications were not observed and later we noted 2 cases of peristomal infections, succesfully managed conservatively. After oncologic surgery we noted 2 (8.69%) pharyngocutaneous fistulas.Conservative care obliterated the fistulas at 6 weeks, maintaining the feeding tube in place. We also compared the results with a group of 27 patients fed through the naso-gastric tube and a group of 20 cases with open gastrotomy-tube prophylactically inserted. The 10 neurological patients had varied conditions but degenerative diseases like motor neuron disease (3 cases" 30%) and multiple sclerosis (2 cases -20%) took the lead we encountered one case of peristomal infection and one case of tube blockage resolved by replacement. We evaluated the nutritional status by controling the weight of these patients before and after PEG placement. A mean weight gain of 3.1 kg(range 1.2 â€" 7) was documented. CONCLUSIONS: PEG is a simple minimmaly invasive procedure performed safely under sedation. It takes a very short time and is virtually free of major complications. The requirements of analgesics are minimal. The refeeding is started early and the tube is well tolerated by the patient. PEG has an important role in the conservative healing of pharyngocutaneous fistula.PEG is the procedure of choice for the neurological patients.It prevents weight loss and aspiration pneumonia in patients with neurogenic dysphagia with a low rate of complications.

Contrast-enhanced ultrasound (CEUS) for the evaluation of focal liver lesions - a prospective multicenter study of its usefulness in clinical practice.

PURPOSE: To assess the value of contrast-enhanced ultrasound (CEUS) for differentiating malignant from benign focal liver lesions (FLLs) and for diagnosing different FLL types. MATERIAL AND METHODS: CEUS performed in 14 Romanian centers was prospectively collected between February 2011 and June 2012. The inclusion criteria were: age > 18 years; patients diagnosed with 1 - 3 de novo FLLs on B-mode ultrasound; reference method (computed tomography (CT), magnetic resonance imaging (MRI) or biopsy) available; patient's informed consent. FLL lesions were characterized during CEUS according to the European Federation of Societies for Ultrasound in Medicine and Biology guidelines. For statistical analysis, indeterminate FLLs at CEUS were rated as false classifications. RESULTS: A total number of 536 cases were included in the final analysis, 344 malignant lesions (64.2 %) and 192 benign lesions (35.8 %). The reference method was: CT/MRI - 379 cases (70.7 %), pathological exam - 150 cases (27.9 %) and aspiration of liver abscesses - 7 cases (1.4 %). CEUS was conclusive in 89.3 % and inconclusive in 10.7 % of cases. To differentiate between malignant and benign FLLs, CEUS had 85.7 % sensitivity, 85.9 % specificity, 91.6 % positive predictive value, 77.1 % negative predictive value and 85.8 % accuracy. The CEUS accuracy for differentiation between malignant and benign liver lesions was similar in tumors with diameter ≤ 2 cm and those with diameter > 2 cm. CONCLUSION: CEUS represents a useful method in clinical practice for differentiating between malignant and benign FLLs detected on standard ultrasonography, and the results of this study are in concordance with previous multicenter studies: DEGUM (Germany) and STIC (France).

The use of gastrostomy procedures in HNC patients.

PURPOSE: To analyse the gastrostomy procedures performed in HNC patients admitted to Coltea Clinical Hospital in order to underline the similarities and differences to the data published worldwide. PATIENTS AND METHODS: Our retrospective study contains 64 HNC cases that met the inclusion criteria between 2008 and 2011. RESULTS AND DISCUSSIONS: The study group presents numerous specific characteristics (a larger number of cases aged over 55 than younger patients; elective use of classic gastrostomy instead of newer techniques; approximately two thirds of the gastrostomies were performed in patients with laryngeal carcinoma; only one third approximately of the cases benefited from prophylactic gastrostomy; etc.). CONCLUSIONS: 22% of the gastrostomies were made after the appearance of a pharyngocutaneous fistula. Therefore we will begin a future prospective study in order to ascertain the value of prophylactic PEG in preventing the appearance of pharyngocutaneous fistulas.

Large charge extraction from metallic multifilamentary Nb3Sn photocathode.

The current density limit for photoemission from metals was measured in an rf photogun to be below 10(9) A/m2. We have achieved 1.6×10(11) A/m2 by photofield emission from a new type of photocathode made from a metallic-composite, multifilamentary Nb3Sn wire driven by a 266 nm picosecond laser pulse and a 2 ns, 50 kV accelerating voltage. This cathode has a micrometer arrayed structure with tens of thousands of Nb/Nb3Sn filaments embedded in a bronze matrix. Our measurements revealed the existence of a new electron emission regime at high laser fluence (100 mJ/cm2). We have extracted stably, and without any surface ablation, up to 4800 pC of charge. This corresponds to 0.9% quantum efficiency, 100 times larger than what is measured from conventional metallic photocathodes. The unexpected large and stable charge extraction cannot be explained by the 3-step model. Thanks to the small emitting area, the measured emittance (0.6 mm·mrad) is low in spite of the high current density and space charge effects. This cathode will be of benefit for many applications based on short and bright electron bunches.

Investigation of six selected bacterial species in endo-periodontal lesions.

AIM: To investigate and determine possible associations of six tested bacteria belonging to 'orange' and 'green' complexes, in endo-periodontal lesions: Parvimonas micra, Fusobacterium nucleatum, Campylobacter rectus, Eubacterium nodatum, Eikenella corrodens and Capnocytophaga sputigena. METHODOLOGY: Forty-six patients presenting with different types of endo-periodontal lesions were investigated. Clinical examinations, periapical radiographs and microbiological sampling from the canal system (endo) and periodontal pockets (perio) were performed. Qualitative and semiquantitative evaluation of bacteria was performed by polymerase chain reaction (PCR) and DNA-DNA hybridization (micro-IDent plus; Hain Lifescience, Germany). RESULTS: Extremely high bacterial loads in endodontic samples were recorded for P. micra, F. nucleatum and C. sputigena, while periodontal samples were often colonized by the same species, plus C. rectus. Significant association was recorded between F. nucleatum-endo and P. micra-endo (P = 0.03, Fisher's exact test). There was marginal evidence of associations between: (i) C. sputigena-endo and C. sputigena-perio (P = 0.06, Fisher's exact test); (ii) P. micra-endo and P. micra-perio (P = 0.05, Fisher's exact test). Sensitivity to percussion was associated with an increased chance of cases with P. micra-endo (P = 0.03, Pearson chi-square test). CONCLUSION: The findings suggest that F. nucleatum, P. micra and C. sputigena may play a role in the pathogenesis of endo-periodontal lesions.

Reconstructive rhinoplasty in cases with basal cell carcinoma of the nose.

Basal cell Carcinoma (BCC) is the most common type of skin cancer, 85% of BCC are located in the head and neck area, of which 30% on the nose. The author present a retrospective study, own experience in surgical treatment in 31 cases with basal cell carcinoma of the nose operated in the period 2005-2011. The age of patients was between 50 to 90 years old. The anatomical site of the nose the most frecvently involved was ala 58%, and followed of the nasal tip 18%, lateral nose wall 12%, dorsum 9% and 3% basalioma terebrans. In all of the cases in this study it was performed the repair of the skin tumour defects of the nose, using varied local skin flaps, septal graft and auricle cartilage free grafts a ndcomposite (skin + cartilage) of conchal and helical rim. Treatment methods were depend on the tumor localization and extension. The best treatment option in BCC of the nose was radical surgical excision whith safety margin of the tumour, followed of reconstructive rhinoplasty.

Nasopharyngeal carcinoma -- analysis of risk factors and immunological markers.

PURPOSE: To analyze the risk factors associated with Nasopharyngeal Carcinoma (NPC)in patients admitted to the ENT Department at Coltea Clinical Hospital in order to assess the similarities and differences as compared to the epidemiological data internationally reported. PATIENTS AND METHODS: This is a retrospective study on 178 cases that met the inclusion criteria from 2003 to 2011. RESULTS AND DISCUSSIONS: There are a number of specific characteristics noticed in our study group: a larger number of cases aged over 60 than younger patients; a linear fall in number of the cases aged between 30-39 years; approximately one third of the cases did not have any of the traditional risk factors; more than 10% of the cases were associated with lymphoma, etc. CONCLUSIONS: The immunologic pattern of our patients presenting undifferentiated NPC is the following: MNF116, CK19, S100, CD34betaE12, Ki67, EBV positive. The rest of the markers were negative (CerB2, EGFR, COX2, p53, CK7, CD117, VGFR, PCNA, L26/CD20, UHCL1, CD15, CD30, VIM, TTF1, CLA, CK17, CEA, LMP, CD79a, EMA).

Tissue-specific dysregulation of hexose-6-phosphate dehydrogenase and glucose-6-phosphate transporter production in db/db mice as a model of type 2 diabetes.

AIMS/HYPOTHESIS: Tissue-specific amplification of glucocorticoid action through 11ß-hydroxysteroid dehydrogenase type 1 (11ß-HSD1) affects the development of the metabolic syndrome. Hexose-6-phosphate dehydrogenase (H6PDH) mediates intracellular NADPH availability for 11ß-HSD1 and depends on the glucose-6-phosphate transporter (G6PT). Little is known about the tissue-specific alterations of H6PDH and G6PT and their contributions to local glucocorticoid action in db/db mice. METHODS: We characterised the role of H6PDH and G6PT in pre-receptor metabolism of glucocorticoids by examining the production of the hepatic 11ß-HSD1-H6PDH-G6PT system in db/db mice. RESULTS: We observed that increased production of hepatic H6PDH in db/db mice was paralleled by upregulation of hepatic G6PT production and responded to elevated circulating levels of corticosterone. Treatment of db/db mice with the glucocorticoid antagonist RU486 markedly reduced production of both H6PDH and 11ß-HSD1 and improved hyperglycaemia and insulin resistance. The reduction of H6PDH and 11ß-HSD1 production by RU486 was accompanied by RU486-induced suppression of hepatic G6pt (also known as Slc37a4) mRNA. Incubation of mouse primary hepatocytes with corticosterone enhanced G6PT and H6PDH production with corresponding activation of 11ß-HSD1 and PEPCK: effects that were blocked by RU486. Knockdown of H6pd by small interfering RNA showed effects comparable with those of RU486 for attenuating the corticosterone-induced H6PDH production and 11ß-HSD1 reductase activity in these intact cells. Addition of the G6PT inhibitor chlorogenic acid to primary hepatocytes suppressed H6PDH production. CONCLUSIONS/INTERPRETATION: These findings suggest that increased hepatic H6PDH and G6PT production contribute to 11ß-HSD1 upregulation of local glucocorticoid action that may be related to the development of type 2 diabetes.

Estrogen receptor alpha polymorphisms: correlation with clinicopathological parameters in breast cancer.

Polymorphisms in estrogen receptor alpha gene (ESR1) have been previously associated with breast cancer risk; however, the results were not fully consistent. Our purpose was to study interactions between common genotypes in ESR1, breast cancer risk and tumor phenotypes. 6 ESR1 single nucleotide polymorphisms (SNPs) were genotyped in 103 breast cancer patients and 90 controls using hybridization probes; the genotypes were correlated with known prognostic factors for breast cancer and 5 years-follow up data. To assess estrogen and progesterone receptors (ER, PR) and HER2/neu expressions, immunohistochemistry was performed. Our results showed that rs3798577 was significantly associated with the risk of breast cancer, the common allele C conferring susceptibility (p-trend=4 x 10(-5)); rs3798577 was also correlated with PR expression (p=0.01), but not with ER expression; rs2228480 (p=0.047) and rs1801132 (p=0.02) were associated with the age at diagnosis; rs1801132 was correlated with hypercholesterolemia (p=0.003) and increased BMI (body mass index) (p=0.01); rs2234693 showed a low significant association (p=0.042) with the tumor grade; rs3798577 was correlated with disease-free survival (p=0.05), allele C conferring increased risk for relapses, but it reached not statistical significance after adjustments. In conclusions, we identified four genotypes significantly correlated with either the risk or some tumor characteristics, suggesting that the main selection criteria of the investigated SNPs (frequency and the position in modulating domains of the gene) are pertinent instruments for establish correlations between the gene structure and the tumor phenotype.

Interaction between estrogens and androgen receptor genes microsatellites, prostate-specific antigen and androgen receptor expressions in breast cancer.

The role of estrogen and androgen receptors signaling in breast cancer is widely accepted, but the interrelations between them are not well understood. It was suggested that PSA could be a marker of endogenous balance between androgens and estrogens. In this context, we intended to investigate the potential of relationship between polymorphic tandem repeats (CAG, TA and CA) in AR (androgen receptor), ERalpha (estrogen receptor alpha) and ERbeta (estrogen receptor beta) genes and the immunoexpression of PSA and AR proteins. We assessed also the possible influences of CAG, TA, and CA variables and other available prognostic factors (ER, PR, AR, HER2/neu, PSA expression, and nodal status) on disease-free survival. We assessed the polymorphic tandem repeats lengths by genotyping, followed by high-resolution denaturing polyacrylamide gel electrophoresis in 163 breast cancers. Immunohistochemistry was performed to assess the expressions of AR, PSA, ER, PR and HER2/neu proteins. Our results showed that PSA was correlated with the length of CA repeats in the 3'-untranslated region of ERbeta, shorter CA repeats being correlated with PSA expression (p=0.03). AR immunoexpression was correlated with CAG repeats on AR gene, higher number of repeats being linked to a higher AR immunoexpression (p=0.04). Performing logistic regression to investigate relationships with prognosis, we observed that PSA immunoexpression (p=0.004), the nodal status (p-<0.001) and marginally, longer TA repeats (p=0.05) were correlated with increased disease-free survival. AR expression presented a low statistical value (p=0.054) in predicting evolution and was not entered into the multivariate regression analysis. Altogether, our findings supports the hypothesis that estrogens, through both alpha and beta-receptors variants are mediating the AR signaling pathway.

Molecular profiling of ADAM12 gene in breast cancers.

ADAMs (a disintegrin and metalloproteinase) family have been associated with the process of proteolytic "shedding" of membrane-associated proteins ectodomain and hence the rapid modulation of key cell signaling pathways in tissues microenvironment. A variety of cytokines, chemokines and growth factors which are initially produced as transmembrane proforms are activated by these sheddase activities. ADAM12 is highly expressed in rapidly growing tissues such as placenta and malignant tumors and it was found as one of the Candidate Cancer Genes in a comprehensive mutational analysis of human breast cancers. Our aim was to determine the gene expression profile of ADAM12 in breast cancers in comparison with normal breast and to correlate their level of expression with the clinical and pathological characteristics of breast cancers. Gene expression of ADAM12 spliced variants (12L and 12S) was evaluated using quantitative reverse-transcription PCR in samples obtained by laser capture microdissection from 38 patients with breast cancers and compared with adjacent healthy breast tissues. Both ADAM12L and 12S expression were significantly up-regulated in breast cancers, while in the normal breast, we found a very low expression. ADAM12L expression was significantly correlated with the histopathological types and, although not statistically significant, ADAM12 both variants were up-regulated in high-grade, highly-proliferative and HER2÷neu positive tumors. From these preliminary results, we found that ADAM12 could be an interesting marker and eventually a therapeutic target for breast cancer.

Mutant HFE genotype leads to significant iron overload in patients with liver diseases from western Romania.

The present study aimed at assessing the frequency of HFE mutations (C282Y, H63D and S65C) in western Romanian patients with liver disease of diverse aetiologies suspected of iron overload. A total of 21 patients, all Romanian residents hospitalized with clinical suspicion of iron overload and liver disease, were assayed for C282Y, H63D and S65C mutations, serum ferritin and viral hepatitis markers. Overall, 9 out of the 21 patients (42.86%) were found to harbour mutations in the HFE gene: 4 homozygotes C282Y (19.0%), 1 compound heterozygote C282Y/H63D (4.8%), 1 single heterozygote C282Y (4.8%), 2 single heterozygotes H63D (9.5%), 1 single heterozygote S65C (4.8%), and 12 wild-type cases (57.1%). Among the subgroup of 10 patients with the most prominent signs of iron overload (hyperferritinaemia and/or hepatocyte iron score > or = 1), without hepatocellular carcinoma, the HFE genotypes were conclusive in 5 cases (50%). They had significantly increased ferritin levels compared to wild-type cases (P = 0.029). The inclusion of iron studies during routine clinical visits, coupled with the availability of HFE genotyping for family and population studies, should facilitate the early detection of hereditary haemochromatosis in Romania.

Differential regulation of prohormone convertase 1/3, prohormone convertase 2 and phosphorylated cyclic-AMP-response element binding protein by short-term and long-term morphine treatment: implications for understanding the "switch" to opiate addiction.

Drug addiction is a state of altered brain reward and self-regulation mediated by both neurotransmitter and hormonal systems. Although an organism's internal system attempts to maintain homeostasis when challenged by exogenous opiates and other drugs of abuse, it eventually fails, resulting in the transition from drug use to drug abuse. We propose that the attempted maintenance of hormonal homeostasis is achieved, in part, through alterations in levels of processing enzymes that control the ratio of active hormone to pro-hormone. Two pro-hormone convertases, PC1/3 and PC2 are believed to be responsible for the activation of many neurohormones and expression of these enzymes is dependent on the presence of a cyclic-AMP response element (CRE) in their promoters. Therefore, we studied the effects of short-term (24-h) and long-term (7-day) morphine treatment on the expression of hypothalamic PC1/3 and PC2 and levels of phosphorylated cyclic-AMP-response element binding protein (P-CREB). While short-term morphine exposure down-regulated, long-term morphine exposure up-regulated P-CREB, PC1/3 and PC2 protein levels in the rat hypothalamus as determined by Western blot analysis. Quantitative immunofluorescence studies confirmed these regulatory actions of morphine in the paraventricular and dorsomedial nucleus of the hypothalamus. Specific radioimmunoassays demonstrated that the increase in PC1/3 and PC2 levels following long-term morphine led to increased TRH biosynthesis as evidence by increased TRH/5.4 kDa C-terminal proTRH-derived peptide ratios in the median eminence. Promoter activity experiments in rat somatomammotrope GH3 cells containing the mu-opioid receptor demonstrated that the CRE(s) in the promoter of PC1/3 and PC2 is required for morphine-induced regulation of PC1/3 and PC2. Our data suggest that the regulation of the prohormone processing system by morphine may lead to alterations in the levels of multiple bioactive hormones and may be a compensatory mechanism whereby the organism tries to restore its homeostatic hormonal milieu. The down-regulation of PC1/3, PC2 and P-CREB by short-term morphine and up-regulation by long-term morphine treatment may be a signal mediating the switch from drug use to drug abuse.

Prostate-specific antigen may serve as a pathological predictor in breast cancer.

PSA (prostate-specific antigen), a serine protease with chymotrypsin-like activity is the most useful tumor marker for prostate cancer screening, diagnosis, prognosis and monitoring. The identification of PSA in normal and tumoral mammary gland was regarded as a curiosity, but the confirmation of PSA expression in the mammary gland by others teams of researchers and the identification of specific mRNA in tumors with PSA immunoexpression initiated new perspectives for studies. The aim of this study was to examine the prevalence of PSA in breast cancers and to evaluate the correlations between PSA expression and some clinicopathological markers. We analyzed the expression of PSA in series of consecutive breast carcinomas by immunohistochemistry and correlated the PSA expression with the histological type and grade, nodal and metastasis status, estrogen receptor (ER), progesterone receptor (PR), androgen receptor (AR) and HER2/neu expression. PSA expression was observed in 44.5% of breast cancers, particularly in lobular types of carcinoma (p<0.0001). In univariate analysis, the expression of PSA was statistically correlated with AR (p<0.0001), PR (p=0.01) and inversely correlated with HER2/neu overexpression (p=0.008) and G3 (p=0.02). PSA did not significantly correlate with ER expression, lymph node and metastasis status. In multivariate analysis, PR was a moderate predictor (p=0.024) but the lobular type (p=0.000), AR (p=0.000), HER2/neu (p=0.002) and G3 (p=0.008) were strong predictors for PSA immunoexpression.

Bcl-2 expression in Hodgkin's lymphoma progression.

INTRODUCTION: Hodgkin's lymphoma study by immunohistochemical expression of Bcl-2 in Hodgkin and Reed-Sternberg cells can precise these cases evolutive way. MATERIAL AND METHODS: Sixty-three cases of classical Hodgkin's disease, hospitalized into the Hematology Department of the County Hospital No. 1 Timisoara, were studied. Histopathological diagnostic was performed using common staining methods, and for revealing the tumoral developments immunohistochemical staining was performed Bcl-2. RESULTS AND DISCUSSION: In our study, the results were noticed a direct relation between the rise of tumoral proliferation index expressions of Bcl-2 and progression of the disease (p < or = 0.001). For I and II stages Bcl-2 expression does not overcome (-/+) category while the III and IV stages, all the cases are situated in (+/-) and (+) categories. No connection we can be noticed between the histological type and Bcl-2 expression although the classic Hodgkin's lymphoma with lymphocyte depletion is considered the most aggressive histological type (p < or = 1). In our study, we found this correlation very important because the main cause of relapses is inadequate staging. In some cases, this staging is difficult; some little lymph nodes could be overlooked because they can be placed in less accessible areas and cannot be evidenced by the most imagistic methods. CONCLUSIONS: All the cases were Bcl-2 expression higher than (+/-) and are staged as I and II stages should be reinvestigated and restaged. This immunohistochemical reaction, although less used in Romania, is very accurate. That is very important because the therapeutically attitude is different in advances stages compared to earlier stages.

Immunohistochemical expression of vascular endothelial growth factor (VEGF) does not correlate with microvessel density in renal cell carcinoma.

The aim of present study was to investigate the relationship between the immunohistochemical expression of vascular endothelial growth factor (VEGF) and microvessel density (MVD) assessed by CD31 and endoglin (CD105) in renal cell carcinoma (RCC). Specimens from 45 cases of RCC. were formalin-fixed, paraffin embedded, and sections were stained with H&E. Additional sections from each case were stained for VEGF, CD31, CD105, and alpha smooth muscle cell actin (SMA). VEGF immunohistochemical expression was estimated as negative (0), weak positive (+1), moderate positive (+2), and intense positive (+3). Microvessel density (MVD) was estimated on 5 hot spots (x400) from each case, and the arithmetic media was the final result. MVD was separately calculated on slides stained with CD31 and CD105. The rate between mature and immature blood vessels was calculated on slides stained with CD31/CD105/SMA. Statistic analysis was performed with SPSS10.0. The immunoreaction for VEGF was positive in epithelial cells of the renal tubules, and occasionally, in endothelial cells. In RCC, tumor cells were positive in 34 from 45 cases (75.5%). 11 cases were negative, 14 were slightly positive (+1), 13 moderate (+2), and 7 intense (+3). No relationship was found between the expression of VEGF and pathological form and nuclear grade, excepting for the chromophilic variant (3 cases, all positive). CD31 was positive in all cases, and CD105 in 39 cases. The mean values of MVD on slides stained for CD31 and CD105 were: 31.68 (range 9.8-60.2)/20.66 (range 4.2-52.8). The rate CD31/SMA positive blood vessels was 1/0.62. VEGF was expressed in 75.5% of 45 cases with RCC, and the mean value of MVD CD31/CD105 was 31.68/20.66. The immunohistochemical expression of VEGF does not correlate with MVD performed on slides stained for both CD31 and endoglin. The majority of blood vessels in the tumor area are of mature type, with perivascular cells positive for SMA.

[Considerations regarding the nosocomial potential of surgical wards]. / Consideratii referitoare la potentialul nosocomial al sectiilor de chirurgie.

PURPOSES: The aim of our study was to determine the prevalence of multiresistant germs with nosocomial potential and their main resistance phenotype and genotype patterns in surgical departments. METHODS: Identification of germs was performed by the API system (BioMerieux France) and susceptibility tests by disk-diffusion tests, (CLSI standards) with automatic reading methods (Osiris-Bio Rad Laboratories). ESBL producing E. coli and Klebsiella pneumoniae strains have been also genotyped. RESULTS: From 190 samples (urines, wound secretions, blood, etc.); we isolated 106 microbial strains with nosocomial potential. 56 (52.83%) from these strains were represented by enterobacteria, 26 (24.52%) by Gram negative non-fermentative rods, and 24 (22.64%) by Gram positive cocci. CONCLUSIONS: We noticed a high prevalence of multidrug resistant germs (ESBL, MRSA, etc). The majority of them were involved in nosocomial surgical site and urinary tract infections.