Epidemiology of Melanoma in Spain: Estimation of Number of Patients With Stage III Disease Eligible for Adjuvant Therapies. / Epidemiología del melanoma en España: estimación de los pacientes con melanoma con estadio III candidatos al tratamiento adyuvante.

BACKGROUND AND OBJECTIVE: Accurate information on the incidence of melanoma by stage and a better understanding of transition between stages are important for determining the burden of disease and assessing the impact of new adjuvant therapies on recurrence and survival. The aim of this study was to estimate the incidence rates of the various stages of melanoma in Spain and to estimate the number of patients with stage III disease who are eligible for adjuvant systemic therapies. MATERIALS AND METHOD: We built an epidemiological model using prospectively collected data from patients diagnosed with de novo or recurrent melanoma between 2012 and 2016 in the melanoma units of 4 public hospitals. RESULTS: The estimated crude incidence rates for stage I and II melanoma were 7 and 2.9 cases per 100,000 person-years, respectively. The corresponding rates for stage III and IV melanoma were 1.9 and 1.3 cases per 100,000 person-years; 25.8% of patients with stage III melanoma were stage IIIA, 47% were stage IIIB, and 27.3% were stage IIIC. The respective estimated incidence rates for recurrent stage III and IV melanoma were 1.1 and 0.9 cases per 100,000 person-years. Overall, 54% of patients with recurrent stage III melanoma had progressed from stage I or II; the other cases corresponded to changes in substage. Of the patients with stage III melanoma, 85% of those with a de novo diagnosis and 80% of those who had relapsed had resectable disease, meaning they were eligible for adjuvant therapy; 47% of these patients had a BRAF mutation. CONCLUSIONS: The above estimates could have a major impact on health care resource planning. Assessing the number of patients with melanoma who are eligible for adjuvant therapies in melanoma could help decision-makers and clinicians anticipate future needs for the management of this disease.

Mitogen-activated protein kinase phosphatase-1 (MKP-1) impairs the response to anti-epidermal growth factor receptor (EGFR) antibody cetuximab in metastatic colorectal cancer patients.

BACKGROUND: The validation of KRAS mutations as a negative marker of response to anti-epidermal growth factor receptor (EGFR) antibodies has meant a seminal advance towards treatment individualisation of colorectal cancer (CRC) patients. However, as a KRAS wild-type status does not guarantee a response to anti-EGFR antibodies, a current challenge is the identification of other biomarkers of response. On the basis of pre-clinical evidence, we hypothesised that mitogen-activated protein kinase phosphatase-1 (MKP-1), a phosphatase that inactivates MAPKs, could be a mediator of resistance to anti-EGFR antibodies. METHODS: Tumour specimens from 48 metastatic CRC patients treated with cetuximab-based chemotherapy were evaluated for KRAS and BRAF mutational status and MKP-1 expression as assessed by immunohistochemistry. RESULTS: As expected, clinical benefit was confined to wild-type KRAS and BRAF patients. Mitogen-activated protein kinase phosphatase-1 was overexpressed in 16 patients (33%) and was not associated with patient baseline clinicopathological characteristics and KRAS mutational status. All patients with BRAF mutations (n=3) had MKP-1 overexpression. Among KRAS wild-type patients, MKP-1 overexpressors had a 7% response rate (RR), whereas patients not overexpressing MKP-1 had a 44% RR (P=0.03). Moreover, median time to progression was significantly longer in MKP-1 non-overexpressing patients (32 vs 13 weeks, P=0.009). CONCLUSION: These results support the concept of MKP-1 as a promising negative marker of response to cetuximab-based treatment in CRC patients with wild-type KRAS.

Trichinella: differential expression of angiogenic factors in macrophages stimulated with antigens from encapsulated and non-encapsulated species.

The newborn larval stage of Trichinella spiralis enters the host striated skeletal muscle cell resulting in the formation of the nurse cell. Vascular Endothelial Growth Factor (VEGF) was detected in cells in the area immediately surrounding the nurse cells. However, no data are available on the antigens involved, the role of other angiogenic factors or the relationship of angiogenesis with Nitric Oxide (NO) production. Using macrophage cell culture we study the effect of different Trichinella L1 antigens from one encapsulated (T. spiralis) and one non-encapsulated (Trichinellapseudospiralis) on the expression of VEGF and basic Fibroblast Growth Factor (FGF2). Also, we investigate the relationship between the production of NO and angiogenic mediators. The results show that encapsulated and non-encapsulated Trichinella species are different in their capacity to stimulate the expression of VEGF and FGF2 from host macrophages. Finally, there is no relationship between angiogenic factors and NO production by T. spiralis antigen.

Serum HER2 extracellular domain predicts an aggressive clinical outcome and biological PSA response in hormone-independent prostate cancer patients treated with docetaxel.

BACKGROUND: Human epidermal growth factor receptor 2 (HER2) overexpression has been linked to hormone-independent prostate cancer (HIPC) progression. Its clinical value and correlation with therapy is not defined. PATIENTS AND METHODS: Patients with HIPC treated with docetaxel (Taxotere) were prospectively tested for serum HER2 extracellular domain (ECD) by immunoanalysis. HER2 was determined by immunohistochemistry and fluorescence in situ hybridization (FISH) in tumor samples. RESULTS: Sixty-nine patients were included. Twenty-four (34.8%) patients had high HER2 ECD (>15 ng/ml). Prostate-specific antigen (PSA) response was 58% for patients with low HER2 ECD and 36% for patients with high HER2 ECD (P = 0.046). HER2 ECD levels were an independent prognostic factor for time to PSA progression [hazard ratio (HR) 2.82; 95% confidence interval (CI) 1.22-6.50; P = 0.015] and overall survival (HR 3.24; 95% CI 1.38-7.59; P = 0.007). Tissue samples from 17 patients were tested for HER2. Patients with negative HER2 tissue expression had lower HER2 ECD levels (median 10.5 +/- 2.7 versus 30.5 +/- 24.8 ng/ml; P = 0.016). FISH was negative in all samples. CONCLUSIONS: High HER2 ECD levels in serum are associated with a worst clinical outcome of HIPC patients treated with docetaxel.

Pharmacological immunomodulation of surgical trauma.

Surgery and accidental trauma induce changes in the immune response, showing a predominant pattern of activation through the Th2 cell pathway to the detriment of Th1 cell pathway activation. Anapsos is a hydrosoluble extract obtained from Polypodium leucotomos. Anapsos has shown immunomodulating effects in vitro. On a rat experimental model (tibia and fibula fracture), cytokines (interleukin [IL]-2, IL-4, IL-6, IL-10, and IL-12) (enzyme-linked immunosorbent assay, ELISA) and cell percentages of CD4, CD8 CD25, CD122, and CD132 (monoclonal antibodies, MoAb) were determined in peripheral blood 7 days before surgery (PRE), 1 day after surgery (1PO), and 7 days after surgery (7PO). On postoperative day 1, rats undergoing fracture show an increase of CD8 percent expression and IL-6 and IL-10 levels, in contrast to rats undergoing fracture plus anapsos treatment. On postoperative day 7, rats undergoing fracture show an increase of IL-6 levels, whereas rats undergoing fracture plus anapsos do not. The IL-12 level decreases on postoperative day 7 in the group with fracture but not in the fracture plus anapsos group. Thus, we conclude that anapsos is able to modulate the immune response after trauma, inhibiting Th2 pathway activation with no effect on Th1 pathway activation. In trauma, Anapsos could prevent the shifting Th1/Th2 balance.

The Sb14-3-3zeta recombinant protein protects against Schistosoma bovis in BALB/c mice.

Schistosoma bovis is a trematode parasite mainly affecting cattle and sheep. Evidences about the arise of drug resistance and the high rates of re-infection of animals in endemic areas have pointed out the need of developing new control tools, e.g., effective vaccines. Schistosomes 14-3-3 proteins have been defined as vaccine candidates against respective infections. We have therefore investigated the protective capacity of the 14-3-3 protein from S. bovis - Sb14zeta - against S. bovis in mice. In addition, we have addressed the influence of the co-administration of four different immunomodulators with the 14-3-3 polypeptide. The values of protection against S. bovis were statistically significant when the Sb14zeta was combined in two independent experiments with the AA0029 (61.0% and 40.31%), AA2829 (49% and 36.3%) and PAL (49% and 40.075%) immunomodulatory molecules. Immune responses from vaccinated animals showed that the highest protection rates do not necessarily match with a dominant Th1-type response.

Progress in the development of Fasciola hepatica vaccine using recombinant fatty acid binding protein with the adjuvant adaptation system ADAD.

Fatty acid binding proteins (FABP) have been designed as a potential vaccine against fasciolosis. In this work, the immunoprophylaxis of the recombinant Fh15 FABP from F. hepatica (Fh15) in adjuvant/immunomodulator ADAD system was evaluated using mice and sheep challenged with F. hepatica. The ADAD system combines the Fh15 antigen with an immunomodulator (hydroalcoholic extract of Polypodium leucotomos; PAL) and/or an adjuvant (saponins of Quillaja saponaria; Qs) in a water/oil emulsion (30/70) with a non-mineral oil (Montanide). All the infected control mice died by 41-48 days post-infection. The mice vaccinated with ADAD only with PAL+Fh15 present a survival rate of 40-50% and those vaccinated with ADAD containing PAL+Qs+Fh15 had a survival rate of 50-62.5%. IgG1 antibodies were lower in surviving mice in comparison with non-surviving mice. The sheep vaccinated with ADAD PAL+Qs+Fh15 showed lower fluke recovery (43%), less hepatic lesions and higher post-infection daily weight gain than F. hepatica infected control animals. Thus, the ADAD system using recombinant fatty acid binding proteins from F. hepatica could be a good option to develop vaccines against F. hepatica.

[Results the application of mitomicyn during endonasal and endocanalicular dacryocystorhinostomy by diode laser]. / Resultados de la aplicación de mitomicina en la dacriocistorrinostomía endonasal y endocanalicular con láser diodo.

AIM OF THE STUDY: To study the effects, secondary effects and security of the intrasurgical application of mitomicyn C during endonasal and endocanalicular dacryocystorhinostomy with diode laser (TLA-ELA DCR). METHODS: We carried out a randomized, prospective, interventional and double blind study in 200 patients: intrasurgical mitomicyn C was applied in 150 of then (0.4 mgr/0.2 ml) for 5 minutes by means of a polivinil acetate dressing over the osteotomy. In other 50 patients MMC was not used we followed up at 24 hours, 10 days and 1, 3 and 6 months after surgery. Endoscopic aspects and possible complications were valued. The results were compared using the Chi-squared test with the Yates correction. We looked for the presence or abscense of secondary effects in the application of mitomicyn C. RESULTS: The average follow up was 15.25 months (range 6 to 21 months). The percentages of alterations for excesive scarring were 21.77% in patients without and MAC 8.03% in the ones MMC. The difference was statisticaly significant (p = 0.02). We did not find secondary effects due to application of mitomcyn. CONCLUSIONS: Intrasurgical application of topical MMC during TLA-ENL DCR reduces the number of pathological findings due to scarring of the surrouding area of the new drainage without secondary effects.

Effect of piroxicam, metamizol, and S-adenosylmethionine in a murine model of experimental trichomoniasis.

Biological effects of piroxicam, metamizol, and S-adenosylmethionine (S-AMET) have been tested in NMRI mice infected intraperitoneally with Trichomonas vaginalis. An intraperitoneal treatment during ten preinfection days with piroxicam (10 mg/Kg/day), or metamizol (275 mg/Kg/day), but not with S-AMET (117 mg/Kg/day) induced a significant decrease of abdominal lesions and mortality, assessed by means of a pathogenicity index. The trichomonicidal activity of piroxicam, metamizol, and S-AMET was tested in vitro at the concentration of 300 microM, but found ineffective. These assays have shown the usefulness of the experimental trichomoniasis model for the study of the immunomodulating activity of synthetic drugs.

Vaccination of mice and sheep with Fh12 FABP from Fasciola hepatica using the new adjuvant/immunomodulator system ADAD.

We evaluate the ability of a Fasciola hepatica FABP native antigen (Fh12) with a new vaccination system called ADAD to protect mice and sheep against an experimental F. hepatica infection. The vaccination protocol consists of a set of two injections. The first injection contains a micelle in which two components are included, saponin from Quillaja saponaria (Qs) and/or Anapsos (A) a Polypodium leucotomos hydroalcoholic extract, both emulsified in a non-mineral oil (Montanide) in a water/oil emulsion (30/70). This is subcutaneously injected to achieve the "adaptation" of the immune system to subsequent stimuli. The second injection contains in addition the Fh12 antigen. Two different experiments were carried out using two mouse strains (BALB/c and CD-1). Mice vaccinated with Qs+A+Fh12 presented a survival rate of 40%, when compared with control groups. Furthermore, we evaluated the efficiency of the vaccination in sheep against an experimental F. hepatica challenge. The vaccinated sheep presented lower fluke recovery (24.5%), number of eggs in bile fluid (58.1%) and faeces (40.3%) than control groups. The recovered flukes were shorter (32.7%), immature (34.0%) and with lower body mass (31.6%) than non-complete vaccinated sheep. Thus, the new ADAD system could be a good alternative for future vaccination experiments against fasciolosis.

[Endonasal and endocanalicular dacryocystorhinostomy by diode laser. Preliminary results]. / Dacriocistorrinostomía endonasal y endocanalicular con láser diodo. Resultados preliminares.

AIM OF THE STUDY: To describe the surgical technique and to evaluate the clinical results after having performed the transcanalicular and endocanalicular dacryocystorhinostomies by diode laser, including the advantages and limits of this technique. METHODS: 34 were performed by diode laser in patients with clinical history of epiphora, with or without mucopurulent secretion, for nasolacrimal duct obstruction. The study was prospective, interventional, non randomized and non comparative. Diode laser was used to realize vaporization of lacrimal sac, osteotomy and vaporization with coagulation of nasal mucosa. The mean of surgical time was 15 minutes (range 7 to 29 minutes). Bicanalicular intubation was performed with a silicone tube and prolene filament for two months in all cases. Postsurgical follow-up was between 4 and 11 months. The degree of epiphora was evaluated by the Munk scale and lacrimal permeability was evaluated by endoscopic functional staining test in all cases. RESULTS: Out of the 34 DCR-EDN+ENC that were performed, 32 cases (94.11%) remain asymptomatic. Two of them (5.88%) required endonasal dacryocystorhinostomies by drilling, because the bony perforation was impossible to achieve by laser fiber. Two cases (5.88%) presented fibrosis and lacrimal and lower canaliculi obstruction, without epiphora because the superior canaliculi was permeable. CONCLUSION: Endonasal and endocanalicular dacryocystorhinostomy technique performed by diode laser is a valid method. It does not cause cutaneous scarring, it decreases thermic canalicular damage, it respects the lacrimal pump, it minimizes pain and bleeding, it needs less surgical time and it has turned into an out-patient procedure with a minimal surgical and postsurgical morbility.

[Transcanalicular dacryocystorhinostomy technique using diode laser]. / Dacriocistorrinostomía transcanalicular con láser diodo.

PURPOSE: To describe the surgical technique and to evaluate the clinical results of carrying out transcanalicular dacryocystorhinostomies using a diode laser (DCR-TC), including the advantages and limitations of this technique. METHODS: 43 DCR-TC were studied and they were analysed using a prospective, interventional, non randomized and non comparative study. We used local and topical anaesthesia in patients with a clinical history of epiphora or dacryocystitis for nasolacrimal obstruction. A diode laser was used to effect a vaporization of the lacrimal sac, osteotomy and vaporization with coagulation of nasal mucosa. The mean duration of surgery was 14 minutes (range 7 to 29 minutes). In all cases, and during a two-month period, bicanalicular intubation was carried out using a silicone tube and prolene filament. Follow-up was between 4 to 38 months. The degree of epiphora was evaluated using the Munk scale and lacrimal permeability was evaluated using Jones I and II tests by direct videoendoscopic control in all cases. RESULTS: 43 DCR-TC were realized, 39 cases are asymptomatic (90.7%). 2 of them (4.65%) had epiphora (degree 2 on the Munk scale) and permeable tract. One case (2.32%) presented lower canaliculi obstruction. One patient showed total osteotomy closing. CONCLUSIONS: Transcanalicular dacryocystorhinostomy carried out using a diode laser is a useful method because it does not cause cutaneous scarring, it hardly produces pain and bleeding, it respects the lacrimal pump, it needs less surgical time, it can be carried out in an out-patient surgery and it generates minimal intra and postsurgical morbility.

Modulation by Polypodium leucotomos extract of cytokine patterns in experimental trichomoniasis model.

The immunomodulating effects of Anapsos, an aqueous hydrosoluble extract obtained from the rhizomes of the fern Polypodium leucotomos, on both pathogenicity and cytokine levels in serum (IFN-gamma/IL-4) were assayed in a Trichomonas vaginalis experimental model (BALB/c mice infected with 10(7) trichomonads and examined at day 15 after infection). Doses of 20 mg/kg/day administered for 10 days before the infection with the parasite induced a decrease of the experimental pathogenicity approximately 10-20% compared to controls. Gross histopathologic changes at abdominal organs and mortality rate, as a consequence of pathogenicity of the protozoa and the immune response of the host, were evaluated. IFN-gamma and IL-4 cytokines were determined on days -5, 0, 5, 10, and 15 postinfection by indirect ELISA. Treatment with PAL before infection modulates and downregulates the IFN-gamma concentration, while anticipates and upregulates the IL-4 level. The assays performed have showed the utility of the murine model of experimental trichomoniasis for the evaluation of immunomodulatory activity of synthetic or natural products.

Effect of Anapsos in a murine model of experimental trichomoniasis.

Immunomodulator effect of Anapsos (Polypodium leukotomas extract) in NMRI (US Naval Medical Research Institute) outbred mice infected by the intraperitoneal route with 10(7) Trichomonas vaginalis has been tested. Gross histopathologic changes in abdominal organs and mortality rate, as a consequence of the pathogenicity of the protozoa and the immune response of the host, were evaluated. Among the different treatment regimes assayed, Anapsos at doses of 20 mg/Kg/day administered for 10 days before infection decreases the parasite pathogenicity index (PI) in the treated animals when compared to those of the untreated control group. The immunosuppressor treatments with azathioprine (100 mg/Kg/day x 1), cyclophosphamide (100 mg/Kg/day x 1), and FK-506 (10 mg/Kg/day x 10) significantly decreased the PI, while an immunostimulant treatment with glycophosphopeptical (13 mg/Kg/day x 10) increased it. These assays have shown the usefulness of the murine model of experimental trichomoniasis for the study of immunomodulator activity of natural or synthetic drugs.

Modulation by Anapsos (Polypodium leucotomos extract) of the antibody responses against the nematode parasite Trichinella spiralis.

The immunomodulant effects of Anapsos, an extract of the naturally occurring fern Polypodium leucotomos was assessed in Balb/c mice immunized with a crude soluble extract (CSE) of Trichinella spiralis L1 larvae. Treatment from day 10 to 1 prior to immunization caused a significant reduction (p < 0.05) in total antibody levels (IgG + M) that was evident from week 2 onwards. Suppression of the IgG1 response was transient, as serum levels were significantly (p < 0. 01) decreased in treated animals during weeks 2 and 3 post-immunization, afterwards increasing to values similar to those in the control group. An opposite pattern was observed in the IgG2a and IgG 2b profiles where, following a brief increase in weeks 3 and 1, respectively, the values fell below those of the control and remained for the whole observation. Anapsos potentiates the IgG3 response against T. spiralis CSE. Deglycosylation of the CSE used in the ELISA assay significantly reduces the IgG3 recognition capacity in both control and treated mice. This treatment did not affect the time-course of an intestinal infection by T. spiralis.