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.

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.

[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.

[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.

[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.

[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.

[Solitary lung metastasis after radical prostatectomy]. / Metástasis pulmonar solitaria tras prostatectomía radical.

Prostate carcinoma is diagnosed in earlier phases of its evolution, but this carcinoma may have an unpredictible evolution. Radical treatment (surgery and radiotherapy) is the best treatment in clinically localized tumors. The biochemical failure over 5 years from the surgery is 20-50% of the patients; the biochemical failure over 10 years from the surgery is less frequent because of prognostic factors from the biologic nature of the tumor. We report a case with biochemical and clinical failure over 10 years from the surgery.

Serotonin upregulates the activity of phagocytosis through 5-HT1A receptors.

1 In this study, we investigated whether serotonin could regulate the in vitro activity of phagocytosis through 5-hydroxytryptamine or serotonin (5-HT(1A)) receptors. 2 Mouse peritoneal macrophages were cultured with serotonin and the activity of phagocytosis was assessed by the uptake of zymosan and latex particles added to the culture media. Specific binding of [(3)H]8-OH-DPAT and immunohistochemistry using an affinity-purified anti-5-HT(1A)-receptor antibody were assayed in the macrophages. In addition, we took advantage of the availability of pharmacological inhibitors of nuclear factor-kappaB (NF-kappaB) to explore its role in the regulation of the 5-HT(1A) receptor. 3 Serotonin increased the in vitro activity of phagocytosis in a dose-dependent manner. The 5-HT(1A) receptor agonist (+/-)-8-hydroxy-2-(di-n-propyl-amino)-tetralin (R(+)-8-OH-DPAT) reproduced these effects. Serotonin- or R(+)-8-OH-DPAT-induced increases in phagocytosis were blocked by the 5-HT(1A) receptor antagonist WAY100635 and the NF-kappaB inhibitor pyrrolidinedithiocarbamate. Moreover, mouse peritoneal macrophages expressed specific binding sites for [(3)H]8-OH-DPAT when cultivated in the presence of zymosan or latex beads. Immunohistochemistry confirmed the expression of the 5-HT(1A) receptor protein in the macrophages. 4 These results show that serotonin can upregulate the activity of peritoneal macrophages through 5-HT(1A) receptors.

[Prognosis factors in pT3 renal carcinoma]. / Factores pronóstico en carcinoma renal pT3.

OBJECTIVES: To identify independent predictors of progression and global survival in patients affected by pT3 renal cell carcinoma. To make risk groups by risk factors. MATERIAL AND METHODS: We evaluated 117 patients with pT3 renal cell carcinoma. 88 was M0 and 29 M1. Most frequent clinical feature: asintomatic patients. 80 males (69%) and 37 females (31%). Mean age 59 (24-82). Median follow up 34 months (mean 44 +/- 39 months). RESULTS: Pathological stage (TNM 1997) was pT3a in 52 patients (43.6%), pT3b 63 patients (53.6%) and pT3c 2 patients. HISTOLOGY: clear cell carcinoma 106 patients (90.6%), papillary 5 patients (4.3%) an dchromophobe 4 patients (3.4%). Nuclear grading according Fuhrman's classification: G1 13 patients, G2 45 patients, G3 32 and G4 12 patients. Size > 4 cm (p = 0.005/p = 0.0019), grade 3-4 (p = 0.006/p = 0.0007), N+ (p = 0.034/p = 0.009) and M+ (p = 0.035/p = 0.042) were independent prognosis factors for progression and global survival of the pT3 renal cell carcinoma. Patients M0 with 0 or 1 risk factor have better global survival tanh patients M0 with 3 or 4 risk factors and patients M1. CONCLUSIONS: Size, grade, N+ and M+ were independent prognosis factors for progression and global survival of the pT3 renal cell carcinoma. Tera are no differencies in global survival between patients M0 with 2 or 3 risk factors and patients M1.

[Usefulness of PET in the diagnosis and response to treatment of extragonadal germinal tumor with atypical presentation]. / Utilidad del PET en el diagnóstico y respuesta al tratamiento en un tumor germinal extragonadal de presentación atípica.

Primary tumors of extragonadal origin are rare, with fewer than 1000 cases described in the literature. Although the exact incidence of EGTs is unknown, clinical data suggest that roughly 3% to 5% of all germ cell tumors. We expose a case report of EGT with unusually clinic presentation. We present our diagnostic and therapeutic experience in this injuries.