Comparative cost analysis of cervical cancer screening programme based on molecular detection of HPV in Spain.

BACKGROUND: HPV cervical cancer screening (CCS) must use validated HPV tests based on the molecular detection of either viral mRNA (Aptima HPV Assay-AHPV) or DNA. AHPV has demonstrated the same cross-sectional and longitudinal sensitivity for the detection of HSIL/CIN2+ lesions but with greater specificity than HPV-DNA tests. The study aimed to estimate the total costs of a CCS with a primary HPV test based on the detection of mRNA compared to DNA in women aged 35-65 years for the National Health System. METHODS: A decision-tree-based model to estimate the cost of the CCS until the first colposcopy was designed based on Spanish CCS guidelines. The total cost (€, 2019) for CCS with AHPV or DNA tests (HC2 and Cobas) was calculated, including HPV test, liquid-based cytology (LBC) and colposcopy, for a population of 7,263,529 women aged 35-65 years (assuming 70% coverage). Clinical inputs derived from a literature review were validated by a multidisciplinary expert panel. Data from head-to-head studies between different HPV tests were selected. RESULTS: The use of AHPV showed reduction of 290,541 (- 35%) and 355,913 (- 40%) LBC compared to HC2 or Cobas, respectively. Furthermore, AHPV avoided 151,699 (- 47%) colposcopies versus HC2 and 151,165 (- 47%) versus Cobas. The total cost of CCS was € 282,747,877 with AHPV, € 322,587,588 with HC2 and € 324,614,490 with Cobas. Therefore, AHPV savings € - 39,839,711 versus HC2 and € - 41,866,613 versus Cobas. CONCLUSIONS: Assuming that 70% of women from 35 to 65 years attend the CCS programme, the cost of screening up to the first colposcopy using AHPV would provide cost savings of up to € 41.9 million versus DNA tests in Spain.

A multistrategic approach in the development of sourdough bread targeted towards blood pressure reduction.

Rising prevalence of hypertension is pushing food industry towards the development of innovative food products with antihypertensive effects. The aim was to study the effect of reduced sodium content and 21% addition of wholemeal wheat sourdough (produced by Lactobacillus brevis CECT 8183 and protease) on proximate composition, γ-aminobutyric acid (GABA) and peptide content of wheat bread. Angiotensin converting enzyme I (ACE) inhibitory and antioxidant activities were also evaluated. Sodium replacement by potassium salt did not affect chemical composition and biological activities of bread. In contrast, GABA and peptides <3 kDa contents in sourdough bread (SDB) were 7 and 3 times higher, respectively, than the observed in control. ACE inhibitory and antioxidant activities of the peptide fraction < 3 kDa from SDB was 1.7 and 2.6-3.0 times higher than control. Therefore, the combination of reduced sodium content with enriched concentrations of bioactive compounds in bread making may provide interesting perspectives for development of innovative breads towards blood pressure reduction.

Changes in bread consumption and 4-year changes in adiposity in Spanish subjects at high cardiovascular risk.

The effects of bread consumption change over time on anthropometric measures have been scarcely studied. We analysed 2213 participants at high risk for CVD from the PREvención con DIeta MEDiterránea (PREDIMED) trial to assess the association between changes in the consumption of bread and weight and waist circumference gain over time. Dietary habits were assessed with validated FFQ at baseline and repeatedly every year during 4 years of follow-up. Using multivariate models to adjust for covariates, long-term weight and waist circumference changes according to quartiles of change in energy-adjusted white and whole-grain bread consumption were calculated. The present results showed that over 4 years, participants in the highest quartile of change in white bread intake gained 0·76 kg more than those in the lowest quartile (P for trend = 0·003) and 1·28 cm more than those in the lowest quartile (P for trend < 0·001). No significant dose-response relationships were observed for change in whole-bread consumption and anthropometric measures. Gaining weight (>2 kg) and gaining waist circumference (>2 cm) during follow-up was not associated with increase in bread consumption, but participants in the highest quartile of changes in white bread intake had a reduction of 33 % in the odds of losing weight (>2 kg) and a reduction of 36 % in the odds of losing waist circumference (>2 cm). The present results suggest that reducing white bread, but not whole-grain bread consumption, within a Mediterranean-style food pattern setting is associated with lower gains in weight and abdominal fat.

Hormone sensitive protocol for metastatic prostate cancer, an update.

To review and update last protocols in hormone sensitive metastatic prostate cancer for improving clinical management in routine. Evidence analysis available about recent updates protocols in hormone sensitive metastatic prostate cancer according to expert panel of clinicians about this field. A nominal consensus group for unify and improve the recommendations to the management of sensitive metastatic prostate cancer patients is currently needed. This document unifies and improve the management of patients with hormone sensitive metastatic prostate cancer, with a methodology that combines data quantitative and qualitative and based on the participation of a broad scientific committee appointed by the Spanish Association of Urology.

The impact of exercise duration and intensity on the release of cardiac biomarkers.

Numerous studies have observed cardiac biomarker release with prolonged exercise. Despite this, we are unsure as to the constituent aspects of any given exercise bout that may be important in promoting cardiac biomarker release. This study examined the influence of exercise duration and intensity on the appearance of cardiac biomarkers. Twenty-one subjects ran for 45, 90 and 180 min at 85% and 95% of their individual anaerobic threshold on six different days randomized. Cardiac troponin I (cTnI) and N-terminal pro-brain natiuretic peptide (NT-proBNP) were assayed from blood samples collected before, 30 min and 3 h post-exercise. NT-proBNP was elevated after all exercise trials (range before: 21-32; range post: 38-67 ng/L). Peak post-exercise concentrations of NT-proBNP were associated with exercise duration (P=0.049), but not exercise intensity (P=0.451). cTnI was elevated after all exercise trials (range before: 0.007-0.011; range post: 0.008-0.021 µg/L). Peak post-exercise concentrations of cTnI were associated with exercise duration (P=0.003) and intensity (P=0.037). Data suggest that while both cTnI and NT-proBNP increased after all exercise trials, the mediating effect of duration influenced both NT-proBNP and cTnI while intensity influenced only cTnI.

Cryptosporidium species and subtype analysis from dairy calves in Spain.

Faecal specimens from 287 diarrhoeic calves younger than 21 days, collected over a 2-year period (2006-2007) from 82 dairy cattle farms in 14 provinces across the north of Spain, were examined for the presence of Cryptosporidium oocysts. Overall, 63 farms (76.8%) and 166 calves (57.8%) tested positive by microscopy. In order to elucidate the genetic diversity, selected positive specimens from 149 calves originating from 61 farms in the 14 provinces were examined by genotyping and subtyping techniques. Cryptosporidium parvum was the only species identified by PCR-RFLP of SSU rDNA from all 149 isolates and sequencing of a subset of 50 isolates, except for 2 specimens that were identified as C. bovis. Sequence analyses of the glycoprotein (GP60) gene revealed that most C. parvum isolates (98%) belonged to the subtype family IIa and 2 isolates were identified as the novel subtype IIdA23G1. Subtype IIaA15G2R1 was the most common and widely distributed (80.3% of the 61 farms), followed by subtype IIaA16G3R1 (14.7%), whereas the remaining IIa subtypes (IIaA16G2R1, IIaA17G2R1, IIaA18G3R1, IIaA19G3R1) were restricted to 1-3 farms. All these C. parvum IIa subtypes have previously been described in human patients, indicating that most isolates from diarrhoeic calves in northern Spain have zoonotic potential.

Virtual quad zygoma implant placement using cone beam computed tomography: sufficiency of malar bone volume, intraosseous implant length, and relationship to the sinus according to the degree of alveolar bone atrophy.

The objective of this study was to investigate the malar bone volume and length that a zygomatic implant can engage, and the relationship to the sinus according to the degree of alveolar bone atrophy. A three-dimensional evaluation was performed using cone beam computed tomography scans from 23 patients with a totally edentulous maxilla; quad zygoma implants were virtually placed. The predictor variable was the amount of malar bone volume and length that a zygomatic implant can engage. The primary outcome variable was the relationship to the sinus according to the degree of alveolar bone atrophy. Other variables were the residual alveolar bone height to the floor of the sinus and the nasal cavity. The mean volume of malar bone engaged in this sample of 92 zygomatic implants was 0.19±0.06cm3. The implant had an extrasinus path in 60.9% of cases, a parasinus path in 25%, and an intrasinus path in 14.1%. The results suggest that the average volume of malar bone engaged by a zygomatic implant is constant regardless of implant position and the degree of alveolar bone atrophy. As alveolar atrophy increases, the trajectory of the implant becomes more parasinus and intrasinus. The examiners were able to find enough bone to adequately distribute the implants in all cases.

Flotillin-1 localization on sporozoites oF Eimeria tenella.

In an attempt to identify parasite surface components involved in the interaction with the host cell, the present research focuses on the rafts of Eimeria tenella that might be involved in the host cell invasion process. To that end, this study was undertaken to investigate the expression of flotillin-1, which is an important component and marker of lipid rafts at the plasma membrane of sporozoites of E. tenella. The expression of this plasma membrane protein was identified by an antibody that specifically reacts with flotillin- and was studied by electron microscopy. Flotillin-1 was found to occur in patches on the surface of E. tenella sporozoites. Immunoblot analysis of the total proteins of the sporozoites showed only 1 band of approximately 48 kDa. This indicates that the antibody exclusively recognized the molecules of flotillin-1 expressed on the surface of E. tenella sporozoites. The presence of flotillin-1 on the cellular membrane of sporozoites predominantly at the apical tip suggests that flotillin-1 belongs to the invasion machinery of E. tenella.

Differences in Hsp70 expression in the sporozoites of the original strain and precocious lines of Eimeria tenella.

The levels of expression of Heat shock protein 70 (Hsp 70) in sporozoites of a wild-type parent strain and 2 precocious lines of Eimeria tenella, were compared to investigate the relationship between the heat shock proteins expressed by the parasite and virulence of the strain. Hsp70 expression was analyzed in sporozoites by immunohistochemical techniques, immunoblot, and flow cytometric analyses. One band of 70 kDa was identified and the variation of the Hsp70 expression levels was quantified by optical densitometric analyses. The results showed a significant gradual decrease in the Hsp70 expression in sporozoites of E. tenella as attenuation progressed, suggesting that the Hsp70 expressed in the excysted sporozoites of E. tenella might be involved in parasite pathogenicity. In addition, the cytoplasmic distribution of the Hsp70, which was observed in the entire sporozoites of the wild strain, was reduced to the anterior portion in the precocious lines.

Comparison of 2 centrifugation procedures in the formalin-ethyl acetate stool concentration technique for the detection of Cryptosporidium oocysts.

A comparison between the standard formalin-ethyl acetate (FEA) stool concentration procedure (centrifugation at 400 g for 2 min) and a modification proposed to increase the recovery of Cryptosporidium oocysts (500 g for 10 min) was performed on 73 formalin-preserved stool specimens known to be positive for Cryptosporidium. False-negative results were obtained with both methods, but the increase in centrifugation time and force resulted in the detection of a higher number of oocysts and positive samples. The modified procedure had a sensitivity significantly higher than the standard method (99% compared to 86%, P = 0.0045).

Eimeria necatrix virus: intracellular localisation of viral particles and proteins.

The presence of the Eimeria necatrix virus was investigated in the following life cycle stages: sporocysts, sporozoites, merozoites, and macrogametes. Electron microscopy revealed virus-like particles (VLPs) in sporozoites, which were purified from sporozoite extracts and used to raise polyclonal antibodies. Viral proteins were identified as RNA polymerase (95 kDa) and the major capsid protein (80 kDa). Polyclonal antibody was used to detect the intracellular localisation of VLPs and proteins. Immunoelectron microscopy and immunohistochemistry identified a viral protein of 95 kDa in all the E. necatrix stages studied, whereas the 80 kDa protein was found only in sporocysts and sporozoites. In addition, no VLPs were found in sporocysts. These results indicate that the synthesis of viral capsid proteins takes place during the early events of sporulation, and is then packaged into novel viruses during the late events. No VLPs were seen and no capsid proteins were found in the merozoites and macrogametes, whereas the 95 kDa RNA polymerase was present in both these stages. In addition, no VLPs or proteins were detected in chicken tissues.

Bioactive sesquiterpenes from Santolina rosmarinifolia subsp. Canescens. A conformational analysis of the germacrane ring.

The hexane extract of aerial parts of Santolina rosmarinifolia subsp. canescens afforded eight new sesquiterpenes in addition to known compounds. Their structures were determined by spectroscopic methods and chemical transformations. The conformational analysis of the germacrane constituents was carried out by spectroscopic methods, including NMR at varying temperature and by molecular mechanics calculations. The antifeedant, antibacterial and antitumoral activity of selected compounds has been tested.

Potential uses and benefits of phytosterols in diet: present situation and future directions.

Over the past decade, the possibility of using phytosterols as ingredients in functional foods has led to numerous research studies in relation to their ability to reduce blood cholesterol. Many different types of carriers have been tested, with good results. The main conclusion is that the effective doses were between 1.5 and 3g/day, leading to reductions between 8% and 15% in LDL-cholesterol. The principal mechanism of action is based on interference with the solubilisation of the cholesterol in the intestinal micelles and, thus, absorption is reduced. Work has also been done on the optimal pattern of administration, and it has been found that ingesting phytosterols in a single dose per day or between meals are equally effective methods. The only side effect is that they can interfere with the absorption of carotenoids, but this can be compensated for in the diet or by adding these compounds in appropriate carriers. It has also been reported that phytosterols have anticancer properties and act as immune system modulators. There are several possible future lines of research: alternative sources with a high phytosterol content must be found, industrial processes must be implemented which minimise their loss, phytosterols must be included in food composition tables, the potential of the different types of phytosterols must be discerned, the genetic bases of their action must be elucidated, synergic effects with other compounds must be studied, side effects must be minimised, and the effects of long-term treatment must be defined precisely.

Detection of Cryptosporidium oocysts in extra-intestinal tissues of sheep and pigs.

Extra-intestinal infections by Cryptosporidium parvum have been detected in pigs and sheep. Detection was carried out by imprints of the mucosa of different organs and viscera in 55 sheep and 57 pigs slaughtered at three abattoirs in Zaragoza (northeast Spain). Imprints were stained by using a modified Ziehl-Neelsen technique. In addition to intestinal infections, cryptosporidial oocysts were found in the gall-bladders of two pigs which were 2 months old, and in some organs of sheep aged 5 days or more, including the gall-bladder (5), mesenteric lymph nodes (2), trachea (7), lung (3) and the uterus of one lamb.

Detection of Blastocystis sp. in pigs in Aragon (Spain).

Infection by Blastocystis sp. has been detected for the first time in pigs in Spain. Detection was carried out by the ethyl acetate-formalin concentration method and direct microscopy. Vacuolated cells of Blastocystis sp. were found in stool samples from 27 of 360 pigs examined (7.5%). The protozoan was present on nine of 17 pig farms studied (53%). Infected pigs ranged from 1 month old to adults, but the organism was most frequent in 1-2 month old (18.4%) and 2-6 month old pigs (15.4%). No correlation was found between Blastocystis sp. infection and occurrence of diarrhoea in pigs.

Cytochemical study of the germinal membrane of the Echinococcus granulosus cyst.

The present cytochemical study was undertaken to provide more information on the localization of enzymatic and glycoconjugates in the germinal membrane of the Echinococcus granulosus cyst. The distinctive distribution of binding sites for two lectins (peanut agglutinin and Dilochos biflorus agglutinin) in the germinal membrane are described. An investigation is made of the distribution and specific activity of adenosine triphosphatase, alkaline phosphatase and acid phosphatase. The results suggest that cells located in the deeper layer of the germinal membrane are intrinsic in the cellular differentiation process. The dissimilarities detected in both the enzymatic activity and the lectin-binding receptors could be associated with metacestode development or degeneration.

Prevalence of intestinal parasites, including Cryptosporidium parvum, in dogs in Zaragoza city, Spain.

Faecal samples from 81 dogs aged between 2 months and 13 years were collected in the small animal clinic (37 domestic dogs) and the animal shelter (44 stray dogs) located in the Faculty of Veterinary Sciences in Zaragoza city (northeast Spain) and screened for the presence of Cryptosporidium oocysts. Faeces were concentrated by the formalin-ethyl acetate method and smears of the sediment were stained by using the modified Ziehl-Neelsen technique. Cryptosporidium parvum oocysts were detected in six dogs (7.4%) aged from 2 months to 6 years. Infection was detected in both domestic (three) and stray (three) dogs and all of them excreted few oocysts (0-1 oocyst per 20 x field). No statistically significant differences in prevalence occurred between dogs younger than 6 months (11.8%) and the older dogs (6.2%). Prevalences were not significantly different between domestic (8.1%) and stray dogs (6.8%). Diarrhoea was recorded in three of the positive dogs (50%), although additional enteric parasites such as oocysts of Isospora spp. were also detected in their faeces. Nevertheless, prevalence was significantly higher in diarrhoeic (30%) versus non-diarrhoeic (4.2%) dogs (P < 0.05). Cryptosporidium was one of the parasites most frequently detected in the dogs surveyed.

Immunohistochemical study of the cyst of Besnoitia besnoiti.

The present study has been undertaken in order to provide information on the molecular structure of the cysts of Besnoitia besnoiti. To that end, immunohistochemical techniques have been used to investigate the expression of several enzymes and proteins implicated in the cellular membrane permeability of bradyzoites. Paraffin and frozen sections, which were obtained from subcutaneous tissue samples taken from naturally infected cattle (coming from northeast Spain), were treated with a panel of antibodies. These were specific for Na(+), K(+)-ATPase, alkaline phosphatase, calmodulin, S100 protein, heat shock proteins, hsp60, and hsp70. Positive-cysts for the said antibodies were found in 23.3% of the cows studied. Bradyzoites showed a positive immunoreaction in every positive cyst with respect to all these antibodies. In addition to the low percentage of positive animals, it is worth noting that positive and unstained cysts were observed in the same tissue section. These results suggest that bradyzoites may pass through both active and dormant metabolic phases.

A method for the sequential study of eimerian chromosomes by light and electron microscopy.

In the present study, the authors describe a simple method to isolate chromosomes from eimerian oocysts and to submit them to sequential study by light and electron microscopy. This method includes a reliable and reproducible technique for transferring eimerian chromosomes from slides to grid that fulfills the essential requirements for generalized use in cytogenetics. In addition, this method overcomes the difficulty of the resistance of protozoan oocysts to disruption and permits the release of intact meiotic chromosomes. The observation by the authors of synaptonemal complexes in meiotic chromosomes of different Eimeria species by applying the above-mentioned method to oocysts revealed its importance to future applications.

Expression of anti-apoptotic factors in cells parasitized by second-generation schizonts of Eimeria tenella and Eimeria necatrix.

Intracellular infections by parasites require a functional anti-apoptotic mechanism for parasite survival within the host cell. The intracellular cycle of Eimeria tenella and Eimeria necatrix in chicken intestinal cells involves the maturation of schizonts within the epithelial cells lining the crypt lumen of the ceca (E. tenella) and jejunum (E. necatrix). After invasion, these cells detach from the epithelial layer and migrate into the underlying connective tissue, where maturation of second-generation schizonts takes place. However, the detached epithelial cells that harbour the parasite and localize in the lamina propia do not undergo apoptosis despite the fact that they are parasitized cells and are located in an inappropriate microenvironment. In this study we consider the hypothesis that E. tenella and E. necatrix may inhibit the host cell apoptosis that accompanies parasite-mediated transformation during late schizogony. To that end, the expression of both NF-kappaB, a transcriptional factor that blocks parasite-induced apoptosis, and bcl-xL, an anti-apoptotic protein induced by NF-kappaB, were studied in the host cell during the maturation of second-generation schizonts. In addition, the expression of the phosphorylated inhibitor of NF-kappaB, p-IkBalpha, was also studied to further confirm NF-kappaB activation. Immunocytochemical techniques, flow cytometric and blott analysis were applied by using polyclonal antibodies that specifically react with bcl-xL, p-IkBalpha, and NF-kappaB to detect these anti-apoptotic proteins in the parasitized cell. Our results offer evidence that both these coccidial species first induce NF-kappaB activation to protect the transformed parasitized cells from apoptosis, allowing the second-generation schizonts to mature, and later, after complete schizonts maturation, cause NF-kappaB inhibition to trigger host cell apoptosis in order to facilitate the escape of merozoites. To determine whether inhibition of the NF-kappaB pathway would induce apoptosis of the host cell, a protease inhibitor (TPCK), which induces apoptosis by mediating inhibition of IkB phosphorylation, was administered to parasitized chickens.