High accuracy of a blood ctDNA-based multimodal test to detect colorectal cancer.

BACKGROUND: Detection of circulating tumor DNA (ctDNA) is a minimally invasive and convenient blood-based screening strategy that may increase effectiveness of colorectal cancer (CRC) screening. PATIENTS AND METHODS: A novel multimodal ctDNA-based blood assay that integrates genomics, epigenomics and fragmentomics, as well as proteomics in a refined version, was tested in blood samples from two cohorts: (i) consecutive fecal immunochemical test (FIT)-positive individuals from the CRC Barcelona stool-based screening program; (ii) patients diagnosed with CRC. Primary endpoint was the performance of the test to detect CRC at different tumor-node-metastasis (TNM) stages. Secondary endpoint was the ability of the test to detect advanced precancerous lesions (advanced adenoma or advanced serrated lesion). RESULTS: A total of 623 blood samples were analyzed in the primary analysis. Sensitivity and specificity of the assay to detect CRC was 93% and 90%, respectively. The sensitivity of CRC detection according to TNM stages was 84% for stage I, 94% for stage II and 96% for stage III (70/73) (P< 0.024). Sensitivity to detect advanced precancerous lesions was 23% with a refined version of the test (including protein and updating bioinformatic thresholding). CONCLUSION: A blood-based multimodal ctDNA assay detected CRC with high accuracy. This minimally invasive, accessible and convenient assay may help to increase the effectiveness of CRC screening.

A quasi-monoenergetic short time duration compact proton source for probing high energy density states of matter.

We report on the development of a highly directional, narrow energy band, short time duration proton beam operating at high repetition rate. The protons are generated with an ultrashort-pulse laser interacting with a solid target and converted to a pencil-like narrow-band beam using a compact magnet-based energy selector. We experimentally demonstrate the production of a proton beam with an energy of 500 keV and energy spread well below 10[Formula: see text], and a pulse duration of 260 ps. The energy loss of this beam is measured in a 2 [Formula: see text]m thick solid Mylar target and found to be in good agreement with the theoretical predictions. The short time duration of the proton pulse makes it particularly well suited for applications involving the probing of highly transient plasma states produced in laser-matter interaction experiments. This proton source is particularly relevant for measurements of the proton stopping power in high energy density plasmas and warm dense matter.

An imbalance between frequency of CD4+CD25+FOXP3+ regulatory T cells and CCR4+ and CCR9+ circulating helper T cells is associated with active perennial allergic conjunctivitis.

Allergic conjunctivitis (AC) is one of the most common eye disorders in ophthalmology. In mice models, it has been suggested that control of allergic conjunctivitis is a delicate balance between Tregs and inflammatory migrating effector cells. Our aim was to evaluate the frequency of Tregs and the frequency of homing receptors expressing cells in peripheral blood mononuclear cells (PBMC) from patients with perennial allergic conjunctivitis (PAC). The analyses of phenotypic markers on CD4+ T cells and both soluble or intracellular cytokines were performed by flow cytometry. CD4+CD25+ cells were 15 times more frequent in PBMC from patients than HC; the vast majority of these CD4+CD25+ cells were FOXP3-, and most of CD4+ T cells were CCR4+ and CCR9+ cells. Upon allergen-stimulation, no significant changes were observed in frequency of Treg; however, an increased frequency of CD4+CCR4+CCR9+ cells, CD4+CD103+ cells and CD4+CD108+ cells with increased IL-5, IL-6, and IL-8 production was observed. These findings suggest an immune dysregulation in PAC, characterized by diminished frequency of Tregs and increased frequency of circulating activated CD4+ T cells; upon allergen-stimulation, these cells were expressing cell-surface molecules related to mucosa homing and were able to trigger an inflammatory microenvironment.

Detection of oocyst-associated toxoplasmosis in swine from southern Chile.

Pork has been traditionally considered an important source of human Toxoplasma gondii infection. Pigs, as other meat-producing animals, can become infected by the ingestion of oocysts that are shed in the environment by infected cats or by the consumption of cysts present in tissues of infected mammals, commonly small rodents. The objective of this study was to investigate the level of T. gondii infection in swine from southern Chile that can be associated with the ingestion of oocysts and therefore exposure to a contaminated environment. A total of 340 serum samples from swine were obtained from three commercial slaughterhouses located in the Araucania and Los Rios Regions from southern Chile. Study animals originated from local farms, mainly small commercial producers, and the meat is sold locally. Overall, 8.8% (30/340) of the samples showed T. gondii-specific IgG antibodies. Of these sero-positive animals, 80% (24/30) were also positive for antibodies specific against the oocyst stage of the parasite, indicating that animals had been infected recently by the ingestion of oocysts. The observed results suggest a high level of environmental contamination with oocysts on the farms of origin. In addition to the food safety problems associated with the consumption of meat from infected animals, the high level of environmental contamination on the farm represents a direct health risk for people living and/or working on these farms. Consequently, there is a need to develop on-farm monitoring programmes and identify risk reduction strategies (food storage, water purification, rodent control and contact with cats) that are appropriate and cost-effective for informal and outdoor type of farms.

Human prophylactic vaccine adjuvants and their determinant role in new vaccine formulations

Adjuvants have been considered for a long time to be an accessory and empirical component of vaccine formulations. However, accumulating evidence of their crucial role in initiating and directing the immune response has increased our awareness of the importance of adjuvant research in the past decade. Nevertheless, the importance of adjuvants still is not fully realized by many researchers working in the vaccine field, who are involved mostly in the search for better target antigens. The choice of a proper adjuvant can be determinant for obtaining the best results for a given vaccine candidate, but it is restricted due to intellectual property and know-how issues. Consequently, in most cases the selected adjuvant continues to be the aluminum salt, which has a record of safety, but predominantly constitutes a delivery system (DS). Ideally, new strategies should combine immune potentiators (IP) and DS by mixing both compounds or by obtaining structures that contain both IP and DS. In addition, the term immune polarizer has been introduced as an essential concept in the vaccine design strategies. Here, we review the theme, with emphasis on the discussion of the few licensed new adjuvants, the need for safe mucosal adjuvants and the adjuvant/immunopotentiating activity of conjugation. A summary of toxicology and regulatory issues will also be discussed, and the Finlay Adjuvant Platform is briefly summarized.

Human prophylactic vaccine adjuvants and their determinant role in new vaccine formulations.

Adjuvants have been considered for a long time to be an accessory and empirical component of vaccine formulations. However, accumulating evidence of their crucial role in initiating and directing the immune response has increased our awareness of the importance of adjuvant research in the past decade. Nevertheless, the importance of adjuvants still is not fully realized by many researchers working in the vaccine field, who are involved mostly in the search for better target antigens. The choice of a proper adjuvant can be determinant for obtaining the best results for a given vaccine candidate, but it is restricted due to intellectual property and know-how issues. Consequently, in most cases the selected adjuvant continues to be the aluminum salt, which has a record of safety, but predominantly constitutes a delivery system (DS). Ideally, new strategies should combine immune potentiators (IP) and DS by mixing both compounds or by obtaining structures that contain both IP and DS. In addition, the term immune polarizer has been introduced as an essential concept in the vaccine design strategies. Here, we review the theme, with emphasis on the discussion of the few licensed new adjuvants, the need for safe mucosal adjuvants and the adjuvant/immunopotentiating activity of conjugation. A summary of toxicology and regulatory issues will also be discussed, and the Finlay Adjuvant Platform is briefly summarized.

Respuesta inmune sistémica y mucosal contra Neisseria meningitidis B inducida por estrategia de vacunación simultànea mucosal y parental

Immunization is one of the most successful and cost-effective health interventions ever. Immunization have been helping to reduce child mortality, improving maternal health and combating infectious diseases. In spite of its, undisputed past success and promising future, however, immunization remains an unfinished agenda because of them inadequate coverage. Several factors have been largely responsible of a difficulty to attain immunization coverage and have been recognized as a problems of current vaccines, such as: the number of dose, excessive use of parenteral route, a small number of adjuvants approve for use in human, higher reactogenicity and unavailability against intracellular pathogens, infected or altered cells and scanty feasibility to combined more than one antigen in the same formulation. For bacterial meningitis WHO estimates that 1,2 million cases occur annually and Neisseria meningitidis is the etiological agent in more than 40 percent of these cases although some meningococcal vaccines are available. To bear in mind these principals problems, a novel protocol for vaccination against N meningitidis called Single Time Vaccination Strategy (SinTimVaS) is proposed. Using female BALB/c mice, we induce systemic and mucosal immune responses against N meningitidis with only one parenteral and one mucosal dose at the same time, employing the Finlay Adjuvants derivate from N meningitidis, AFPL1 and AFCo1, respectively. In conclusion, SinTimVaS could increase the vaccination coverage and reduce the time-cost of vaccine campaigns, adding the possibility to increase the herd immunity by mucosal specific response induction(AU)

Enfoques mucosales en vacunologia de Neisseria

Meningococcal B strains accounts for some 72 percent and 28 percent of meningococcal diseases in infants and toddlers in Europe and the USA, respectively. Nevertheless, meningococcal diseases are rare in Cuba owing to the wide spread program on antimeningococcal vaccination in the country. Finlay Institute is one of the pioneering organizations in Neisseria Vaccinology mainly by its contribution to N. meningitidis serogroup B outer membrane-based bivalent vaccine, VA-MENGOC-BC™. This vaccine was given intramuscularly in more than 60 million doses corresponding 10,7 millions of them to Cuban young adults, children, and infants. However, most dangerous or commensally Neisseria strains enter and establish in the mucosa, where the secretory (S) IgA is the main specific guardian and is mainly induced by mucosal routes. However, few mucosal vaccines exist principally due to the absent of mucosal adjuvants. We develop a Finlay Adjuvant (AF) platform based in outer membrane vesicles (Proteoliposome, PL) and its derivate Cochleate (Co). AFPL1 derived from serogroup B N meningitidis is a potent Th1/CTL driving parenteral adjuvant. AFCo1 is a potent mucosal adjuvant. Therefore, we sought to go deeper in the possible mucosal cross recognition between N. meningitidis serogroups and Neisseria species and explore a concurrent mucosal and parenteral immunization strategy (SinTimVaS) in order to develop suitable mucosal vaccines. Experiments were conducted in Balb/c or C57Bl6 mice with mucosal and systemic immunization using AFCo1 and AFPL1. Human sera and saliva were also analyzed for cross cognition. Mucosal cross recognition at SIgA level in human saliva between N. meningitidis serogroups B, A, C, Y, and W135 were observed. This SIgA cross recognition response was also observed between pathogenic (N meningitidis serogroup B, N gonorrhoeae) and non-pathogenic strains (N flava, N lactamica). The possible influence of meningococcal vaccination ...(AU)

New vaccines require potent adjuvants like AFPL1 and AFCo1.

Neisseria meningitidis B proteoliposome (AFPL1 when used as adjuvant) and its derivative-Cochleate (AFCo1) contain immunopotentiating and immunomodulating properties and delivery system capacities required for a good adjuvant. Additionally, they contain meningococcal protective antigens and permit packaging of other antigens and pathogen-associated molecular patterns (PAMP). Consequently, we hypothesized that they would function as good vaccine adjuvants for their own antigens and also for non-related antigens. AFPL1 is a detergent-extracted outer membrane vesicle of N. meningitidis B transformed into AFCo1 in calcium environment. Both are produced at Finlay Institute under good manufacture practices (GMP) conditions. We show their exceptional characteristics: combining in the same structure, the potentiator activity, polarizing agents and delivery system capacities; presenting multimeric protein copies; containing multiprotein composition and multi and synergistic PAMP components; acting with incorporated or co-administrated antigens; inducing type I IFN-gamma and IL-12 cytokines suggesting the stimulation of human plasmocytoid precursor and conventional dendritic cells, respectively, inducing a preferential Th1 immune response with TCD4(+), TCD8(+), cross-presentation and cytotoxic T-lymphocyte (CTL) in vivo responses; and functioning by parenteral and mucosal routes. AFPL1-AFCo1 protective protein constitutions permit per se their function as a vaccine. In addition to Phase IV Men BC vaccine, AFPL1 has ended the preclinical stage in an allergy vaccine and is concluding the preclinical stage of a nasal meningococcal vaccine. In conclusion, AFPL1 and AFCo1 induced signal 1, 2 and 3 polarizing to a Th1 (including CTL) response when they acted directly as vaccines or were used as adjuvants with incorporated or co-administered antigens by parenteral or mucosal routes. Both are very promising adjuvants.

[Polyarthritis secondary to intravesical BCG instillation, a case report]. / Poliartritis secundaria a instilaciones intravesicales de BCG, a propósito de un caso.

Immunotherapy with intravesical instillation of Bacillus Calmette is the best complementary after TUR treatment against T1 grade 2- 3 and CIS bladder cancer. However, this therapy is associated with several side- effects, incluiding joint symptoms. In this article we describe a case of polyarthritis associated with intravesical instillation, a rare complication. We review the clinical features, the pathogenic mechanisms and treatments described previously in the medical literature. Finally, we expose our brief experience and our final result.

[Local relapse and single site of metastatic involvement of renal tumour. Prognostic factors and survival]. / Recidiva local y afectación metastásica única de tumor renal. Factores pronósticos y supervivencia.

OBJECTIVE: To evaluate the clinical and pathological characteristics and survival in patients surgically treated for renal tumours that had local recurrence or metastasis to a single site. MATERIAL AND METHODS: A retrospective study of 321 nephrectomies, evaluating the clinical and pathological variables in patients having local recurrence or metastasis to a single site, and who were treated surgically. Study and comparison of survival in the different groups. RESULTS: The only factor found to have an independent influence on local recurrence is pathological stage. Local recurrence and the presence of metastasis to a single site have similar survival rates, both being statistically worse than in patients without metastasis at diagnosis, but better than in those having metastasis at diagnosis. CONCLUSIONS: The presence of local recurrence has the same prognosis as a single excisable metastatic site, the prognosis being better than those initially with metastasis subjected to nephrectomy before receiving systemic treatment.

[Pheochromocytoma with thrombus in cava]. / Feocromocitoma con trombo en cava.

Pheochromocytoma, a paraganglioma of suprarenal location, is a catecholamine-secreting chromaffin cell tumour. Spread of these tumours to the vena cava is rare and the thrombus only reaches the right atrium in exceptional cases. We present the case of a patient who, without previous symptomatology, presented with a clinical picture of multiorganic dysfunction with primary manifestation of a suprarenal tumour with vascular spread to the right atrium affecting the right suprahepatic vein.

[Evaluation and prognostic of cystic renal tumors]. / Valoración y pronóstico de los tumores renales quísticos.

OBJECTIVE: To evaluate the clinical and pathological characteristics of cystic renal tumors in our center. MATERIAL AND METHODS: A retrospective review of 239 nephrectomies is performed comparing the clinical and pathological variables of cystic tumors with those of solid renal tumors. Survival outcomes are analyzed in both groups. RESULTS: Our experience shows that cystic renal tumors behave like solid renal tumors, with no differences in survival shown. The variables studied show statistically significant differences in histological grade and number of tumors, with cystic tumors having a lower histological grade and being more often multiple in number. CONCLUSIONS: The behavior of cystic renal tumors is no different than that of solid renal tumors.

Recidiva local y afectación metastásica única de tumor renal. Factores pronósticos y supervivencia / Local relapse and single site of metastatic involvement of renal tumour. Prognostic factors and survival

Objetivo: Valorar las características clínicas y patológicas y la supervivencia de los tumores renales que presentaron recidiva local o metástasis única y fueron tratadas quirúrgicamente. Material y método: Estudio retrospectivo de 321 nefrectomías valorando las variables clínicas y patológicas de aquellos pacientes que presentaron recidiva local o metástasis única y fueron tratados quirúrgicamente. Estudio y comparación de la supervivencia en los diferentes grupos. Resultados: El único factor influyente de forma independiente en la presentación de recidivalocal es el estadio patológico. La recidiva local y la presencia de metástasis única tienen una supervivencia similar, estadísticamente peor que el resto de pacientes no metastásicos al diagnóstico, pero mejor que los pacientes con presencia de metástasis al diagnóstico. Conclusiones: La presencia de recidiva local tiene el mismo pronóstico que la progresión en forma de una única metástasis resecable, pero mejor pronóstico que los pacientes inicialmente metastásicos a los que se realiza nefrectomía previa al tratamiento sistémico (AU)

Objective: To evaluate the clinical and pathological characteristics and survival in patients surgically treated for renal tumours that had local recurrence or metastasis to a single site. Material and Methods: A retrospective study of 321 nephrectomies, evaluating the clinical and pathological variables in patients having local recurrence or metastasis to a single site, and who were treated surgically. Study and comparison of survival in the different groups. Results: The only factor found to have an independent influence on local recurrence is pathological stage. Local recurrence and the presence of metastasis to a single site have similar survival rates, both being statistically worse than in patients without metastasis at diagnosis, but better than in those having metastasis at diagnosis. Conclusions: The presence of local recurrence has the same prognosis as a single excisable metastatic site, the prognosis being better than those initially with metastasis subjected to nephrectomy before receiving systemic treatment (AU)

Feocromocitoma con trombo en cava / Pheochromocytoma with thrombus in cava

El feocromocitoma, paraganglioma de localización suprarrenal, es un tumor cromafín secretor de catecolaminas. La extensión de éstos tumores a vena cava es rara y que el trombo alcance la aurícula derecha es excepcional. Presentamos el caso de una paciente que, sin clínica previa, presentó un cuadro de disfunción multiorgánica como primera manifestación de un tumor suprarrenal con extensión vascular hasta la aurícula derecha y afectación de la vena suprahepática derecha (AU)

Pheochromocytoma, a paraganglioma of suprarenal location, is a catecholamine-secreting chromaffin cell tumour. Spread of these tumours to the vena cava is rare and the thrombus only reaches the right atrium in exceptional cases. We present the case of a patient who, without previous symptomatology, presented with a clinical picture of multiorganic dysfunction with primary manifestation of a suprarenal tumour with vascular spread to the right atrium affecting the right suprahepatic vein (AU)

[Incidentally detected renal cancer. Prognostic factors]. / Tumores renales incidentales. Factores pronósticos.

OBJECTIVE: To study the clinical and pathological characteristics of incidental renal tumors treated in our center. MATERIAL AND METHODS: A retrospective review is conducted of 318 nephrectomies comparing the clinico-pathological variables of renal tumors diagnosed incidentally with those of symptomatic renal tumors. The factors influencing disease-free survival are analyzed in both groups. RESULTS: In our experience, although incidental renal tumors presented better survival than symptomatic ones owing to their better pathological state and tumor grade, incidental diagnosis was not an independent influencing factor in the multivariate study. Only when patients were studied who did not present metastases on diagnosis did incidental diagnosis become an influencing factor very close to statistical significance. CONCLUSIONS: Incidental diagnosis is not an independent prognostic factor.

[Follicular carcinoma of the thyroid manifested initially as asymptomatic primary renal neoplasm]. / Masa renal asintomática como primera manifestación de carcinoma folicular de tiroides.

Metastases in the kidney are rare, evenmore if primary source is thyroid. We report the tenth case of metastases in the kidney from thyroid, and it is the first to be follicular type and absolutely asymptom. Sonography and computerized tomography with suspicion of renal tumour are showed in a asymtom female 75 years old. Left partial nephrectomy was perfomed, initially it has been pathologically diagnosed as renal clear cells tumour, however the definitive pathologic report showed follicular tumour of thyroid. Local and systemic stage was discovered with complementary techniques. Sources of metastases in kidney and diagnoses techniques are discussed.

[Mutations of the androgen receptor gene in patients with clinically localized adenocarcinoma of the prostate]. / Mutaciones del gen del receptor de andrógenos en pacientes con adenocarcinoma de próstata clínicamente localizado.

UNLABELLED: The aim of this study was to detect mutations in the human androgen receptor gene in radical prostatectomy specimens. MATERIAL AND METHODS: The genomic sequence was realized in 67 radical prostatectomy specimens. The mean age was 64 years old. The PSA median was 15 ng/ml. TNM 1997: 34.3% were T1 and 65.7% T2. Genomic sequence: 1. Radical prostatectomy specimens desparaffitation. 2. Extraction of the DNA 3. DNA amplification. 4. Automatic genome sequence. 5. Comparison with Gene-Bank. RESULTS: 16.7% of the specimens were mutated. The most frequent mutation was the punctual mutation. The exon most frequent mutated was exon 1.

[Sexual behaviour and use of condoms by adolescents in our environment]. / Comportamientos sexuales y uso de preservativos en adolescentes de nuestro entorno.

OBJECTIVE: To identify sexual behaviour in adolescents. DESIGN: Transversal, descriptive study, by means of an anonymous structured questionnaire. SETTING: Urban secondary school. PARTICIPANTS: Simple random sample of 84 students (4th year of ESO--c. 15/16 years old). MAIN MEASUREMENTS: Personal details, sexual practices, use of condoms. RESULTS: 61.9% were girls. Average age was 16.4 (95% CI, 16.2-16.6). They had a girl/boy-friend or casual partner (33.7%) and stable partner (60.7%). 82.2% had total or sufficient trust. Boys' first masturbation was between 14 and 16 (46.8%). 100% had done so by the age of 16. 72.3% of girls had never masturbated at this age (P<.001).Boys' first kiss on the lips occurred between 14 and 16 (46.8%); 100% had done so by 16. 60.7% of girls had done so between 14 and 16, and 92.2% by 16.30.1% of boys had had their first experience of coitus by the age of the questionnaire, whereas 22.5% of girls had. Both sexes used a condom on 71.4% of occasions.38.1% had sex with penetration, 46.2% masturbated. 22.2% had had 2 sexual partners, 8.9% had had 3, and 40.0% had had 4 or more. Condoms were used in 83.3% of relationships with penetration. CONCLUSIONS: This is a sexually active population, susceptible to sexually transmitted diseases (STD) and unwanted pregnancies, as it does not use condoms in every relationship involving penetration. Health education activities need to be encouraged.

[Female incontinence: urodynamic evaluation]. / Incontinencia femenina: evaluación urodinámica.

OBJECTIVE: To evaluate the functional studies in women with stress urinary incontinence. METHODS: Emphasis is placed on comprehensive understanding of stress urinary incontinence in women as well as Urodynamic techniques and findings which apply to women with this condition. RESULTS: Although prospective randomized controlled trials to evaluate the clinical efficacy, cost-effectiveness and effect on quality of life of a pre-operative Urodynamic assessment are necessary, to date only Urodynamic Investigation (Non-invasive uroflowmetry, filling cystometrogram, valsalva leak point pressure, pressure-flow studies and urethral profile) provide enough information for treatment decisions and prognosis in cases of female urinary incontinence.