Colonic adenosquamous carcinoma and mucinous adenocarcinoma with microsatellite instability.

A 43-year-old man presented with two-month history of fatigue, weakness, paleness, rectal bleeding, sweating, and weight loss of 10 kg in the past one month. A complete blood count revealed anaemia. The patient underwent a right hemicolectomy. The microscopic examination revealed an adenosquamous carcinoma associated with a mucinous adenocarcinoma in a patient with microsatellite instability due to loss of MLH1 and PMS2 expression and retention of MSH2 and MSH6 expression in both the squamous and glandular components. We also observed an atypical immunohistochemical phenotype in the adenocarcinoma component showing CK7 expression and reduced CK20 and CDX2 expression.

Role of primary care in the follow-up of patients with obstructive sleep apnoea undergoing CPAP treatment: a randomised controlled trial.

OBJECTIVE: To evaluate whether follow-up of patients with obstructive sleep apnoea (OSA) undergoing CPAP treatment could be performed in primary care (PC) settings. DESIGN: Non-inferiority, randomised, prospective controlled study. SETTINGS: Sleep unit (SU) at the University Hospital and in 8 PC units in Lleida, Spain. PARTICIPANTS: Patients with OSA were randomised to be followed up at the SU or PC units over a 6-month period. MAIN OUTCOMES MEASURED: The primary outcome was CPAP compliance at 6 months. The secondary outcomes were Epworth Sleep Scale (ESS) score, EuroQoL, patient satisfaction, body mass index (BMI), blood pressure and cost-effectiveness. RESULTS: We included 101 patients in PC ((mean±SD) apnoea-hypopnoea index (AHI) 50.8±22.9/h, age 56.2±11 years, 74% male) and 109 in the SU (AHI 51.4±24.4/h, age 55.8±11 years, 77% male)). The CPAP compliance was (mean (95% CI) 4.94 (4.47 to 5.5) vs 5.23 (4.79 to 5.66) h, p=0.18) in PC and SU groups, respectively. In the SU group, there were greater improvements in ESS scores (mean change 1.79, 95% CI +0.05 to +3.53, p=0.04) and patient satisfaction (-1.49, 95% CI -2.22 to -0.76); there was a significant mean difference in BMI between the groups (0.57, 95% CI +0.01 to +1.13, p=0.04). In the PC setting, there was a cost saving of 60%, with similar effectiveness, as well as a decrease in systolic blood pressure (-5.32; 95% CI -10.91 to +0.28, p=0.06). CONCLUSIONS: For patients with OSA, treatment provided in a PC setting did not result in worse CPAP compliance compared with a specialist model and was shown to be a cost-effective alternative. TRIAL REGISTRATION NUMBER: Clinical Trials NCT01918449.

Objective Bayesian meta-analysis for sparse discrete data.

This paper presents a Bayesian model for meta-analysis of sparse discrete binomial data, which are out of the scope of the usual hierarchical normal random-effect models. Treatment effectiveness data are often of this type. The crucial linking distribution between the effectiveness conditional on the healthcare center and the unconditional effectiveness is constructed from specific bivariate classes of distributions with given marginals. This assures coherency between the marginal and conditional prior distributions utilized in the analysis. Further, we impose a bivariate class of priors that is able to accommodate a wide range of heterogeneity degrees between the multicenter clinical trials involved. Applications to real multicenter data are given and compared with previous meta-analysis.

Bayesian heterogeneity in a meta-analysis with two studies and binary data.

The meta-analysis of two trials is valuable in many practical situations, such as studies of rare and/or orphan diseases focussed on a single intervention. In this context, additional concerns, like small sample size and/or heterogeneity in the results obtained, might make standard frequentist and Bayesian techniques inappropriate. In a meta-analysis, moreover, the presence of between-sample heterogeneity adds model uncertainty, which must be taken into consideration when drawing inferences. We suggest that the most appropriate way to measure this heterogeneity is by clustering the samples and then determining the posterior probability of the cluster models. The meta-inference is obtained as a mixture of all the meta-inferences for the cluster models, where the mixing distribution is the posterior model probability. We present a simple two-component form of Bayesian model averaging that is unaffected by characteristics such as small study size or zero-cell counts, and which is capable of incorporating uncertainties into the estimation process. Illustrative examples are given and analysed, using real sparse binomial data.

Pla 1 1 and Ole e 1 pollen allergens share common epitopes and similar ultrastructural localization.

BACKGROUND: English plantain (Plantago lanceolata L.) and olive (Olea europaea L.) pollens are important causes of pollinosis in large areas of North America, Australia, and the Mediterranean basin. The major pollen allergens of both plants, Pla I 1 and Ole e 1, share 38.7% of their amino acid sequences. OBJECTIVE: To analyze putative cross-reactivity between these 2 proteins. METHODS: Several antibodies and patients' sera were used in immunoblot and immunocytochemistry experiments. RESULTS: Two anti-Pla I 1 antibodies were able to bind to 3 polypeptides from olive pollen protein extracts, which correspond to the 3 glycosylation isoforms of Ole e 1 (18-22 kDa) previously described. Moreover, Pla I 1 protein was found in the cytoplasm of both the vegetative and the generative cells of P lanceolata mature pollen. On olive pollen sections, these anti-Pla I 1 antibodies displayed significant labeling in the cytoplasm of the vegetative cell and in both the exine and the material adhering to this outer layer of the pollen wall. In addition, the anti-Ole e 1 antibody 10H1 was found to cross-react with proteins of similar masses (16-20 kDa) to Pla I 1 variants. In Plantago pollen sections, the 10H1 antibody recognized proteins located in the cytoplasm of both the vegetative and generative cells. Cross-reaction was confirmed using sera from patients allergic to either plant pollen. CONCLUSION: Both allergens share common epitopes, which can be cross-recognized by different antibodies and sera from different patients, although this antigenic similarity seems to have little clinical relevance.

[Sacral root neuromodulation as treatment for fecal incontinence. Preliminary results]. / Neuromodulación de raíces sacras como tratamiento de la incontinencia fecal. Resultados preliminares.

We present our initial experience in the treatment of fecal incontinence (FI) with sacral root neuromodulation (SRN) by reporting the results of a prospective study with 26 patients where baseline Wexner-Cleveland scale scores and ability to delay defecation were compared to results after one year with SRN. The initial study of patients included history taking, general examination, anal ultrasonography, and manometry, and a three-week diary of continence and quality of life specific for FI was used. Before SRN the mean baseline Wexner-Cleveland score was 15.00 +/- 1.81, and 62.50% of patients could only delay defecation for less than a minute. After a year with NRS the mean Wexner-Cleveland score was 4.87 +/- 2.54 (p = 0.0031), and 75.01% of patients could delay defecation above fifteen minutes (p = 0.0018). We also describe the surgical technique and its indications, and finally review the various therapeutical options for FI and show our algorithm for this condition. SRN is an effective technique for the treatment of FI in properly selected patients with no response to medical therapies (including biofeedback) or anatomic correction (sphincteroplasty), with efficacy, little morbidity, and a short hospital stay.

Studies on the relationship between structure and IgE-binding ability of Parietaria judaica allergen I.

The main allergen of Parietaria judaica pollen, Par j I, is a glycopolypeptide with mol. wt about 10,000. It shows a considerable charge heterogeneity which is mostly due to the carbohydrate prosthetic groups, since treatment with trifluoromethanesulfonic acid yielded a deglycosylated protein with 8500 mol. wt that displayed only a few bands on IEF, in a narrow pH-region around 5.0. Deglycosylated Par j I exhibited a specific allergenic activity slightly lower than that of native Par j I; however, no allergenic determinants should be located on the sugar moiety since both native and deglycosylated Par j I inhibited up to a similar extent the binding of specific human IgE to P. judaica-coated wells in ELISA. The decrease of specific allergenic activity following deglycosylation could be ascribed to conformational changes evidenced by CD experiments. On the other hand, fluorescence spectroscopy showed that Par j I bears unidentified yellow-brown chromophores strongly linked to the polypeptide chain. These chromophores were not removed by TFMS treatment. Finally, reduction and alkylation caused the complete loss of allergenic activity, showing that disulphide bridges are essential for the IgE-binding ability of Par j I.

HPLC purification of the main allergen of Parietaria judaica pollen.

A two-step purification procedure of Par j I from the whole Parietaria judaica pollen extract is described. The first step consisted of gel filtration HPLC using a TSKG 3000 SW column, and 0.1% trifluoroacetic acid as the eluant. By this method, proteins were separated from the highly colored material present in the extract. Then, Par j I-containing fractions were chromatographed on a reversed-phase HPLC column (Vydac C4) using an acetonitrile gradient. This second step yielded pure Par j I as assessed by SDS-PAGE and CIE. Previously reported microheterogeneity was still observed, but amino acid analysis of various RP-HPLC fractions suggested that the heterogeneity of Par j I might not be due to changes in its primary structure. Allergenic activity of Par j I was shown to be retained after the purification procedure by several immunochemical techniques.

Purification of Par j I, the major allergen of Parietaria judaica pollen.

A component of Parietaria judaica pollen extract, previously identified as the major allergen, then reported as Pj10 and hereafter denominated Par j I has been isolated by a combination of 65% ammonium sulphate salt precipitation and gel filtration and an Ultrogel AcA54 column. The purified allergen appeared essentially homogeneous on gel filtration HPLC. The mol. wt of Par j I was estimated by electrophoretic and chromatographic techniques. All results gave values in a range from 13 K to 25 K. Analysis in SDS-PAGE under reducing conditions revealed a broad band corresponding to a mol. wt of 10 K, which retained allergenicity when tested with a patients serum pool. CIE and CRIE patterns of the isolated Par j I displayed the two precipitating lines already reported as those exhibiting the highest IgE-binding ability. Par j I showed a specific allergenic activity about 10-fold higher than that of the whole extract and was demonstrated to be the major allergen of Parietaria judaica as assessed in 25 sensitive human sera.

Studies on the biochemical structure of the major cat allergen Felis domesticus I.

The major cat allergen, Fel d I, was purified to homogeneity from cat dander extract by sequential mAb affinity chromatography and HPLC size exclusion. The purity and allergenic activity of the preparation was demonstrated by different techniques such as HPLC, RAST inhibition, skin prick tests and CIE/CRIE. Fel d I showed a mol. wt of about 35,000 by HPLC gel filtration and of 18,000 by SDS-PAGE, confirming that it is a non-covalently linked dimer. However, SDS-PAGE analysis under reducing conditions as well as labelling experiments with 14C-iodoacetamide of 2-ME-reduced Fel d I showed that each mol. wt 18,000 monomer is comprised of two covalently S-S bound polypeptides with apparent mol. wt. of 4000 (alpha-chain) and 14,000 (beta-chain). Reduction and alkylation of Fel d I obliterated most of its allergenic activity, as determined by RAST inhibition and immunoblotting, suggesting that most of the IgE-binding sites are conformational. On the other hand, treatment of Fel d I by N-glycanase under reducing and non-reducing conditions indicated the presence of N-linked oligosaccharides in the beta-chain. Carbohydrate analysis data of the whole Fel d I molecule showed the presence of a relatively high carbohydrate content (approximately 20%). RAST inhibition experiments of native and deglycosilated allergen suggest that most IgE epitopes are located in the protein moiety of the molecule. However, the deglycosilated allergen showed a 2-4 fold reduction in its inhibition capacity of RAST as compared to the native allergen, suggesting that carbohydrates could have some role in keeping the active conformation of those epitopes. The N-terminal amino acid sequence of the beta-chain (20 residues) and most of the alpha-chain (40 residues) were determined. Both chain sequences showed no homology with other known protein sequences.

Isolation by mAb based affinity chromatography of two Par j I isoallergens. Comparison of their physicochemical, immunochemical and allergenic properties.

We report the identification and separation of two isoallergen components of Par j I, the major allergen from Parietaria judaica pollen. First, electrophoretic conditions for consistently separating both isoforms in an SDS-PAGE system were established, and mol. wt values of 13,000 (isoallergen IA) and 10,500 (isoallergen IB) were estimated. Immunoblot, after SDS-PAGE experiments, with individual P. judaica-sensitive human sera revealed a slightly different IgE-binding pattern for each isoallergen. Four anti-Par j I mAbs were obtained from BALB/c mice immunized with a purified Par j I preparation comprising IA and IB isoallergens. Three mAbs were directed to an epitope shared by both isoallergens, and the fourth one recognized specifically one epitope on Par j IB. Dot-blot experiments with the deglycosylated allergen showed that the mAbs did not recognize the carbohydrate prosthetic group of the molecules. Affinity chromatography using the mAbs allowed the separation of the isoallergens that retained their IgE-binding ability after the purification process. Amino acid composition analyses and partial N-terminal sequencing demonstrated an extensive homology and also the existence of some structural differences between Par j I isoallergens, which is in agreement with the high, but not complete, cross-reactivity observed in competition ELISA experiments. Finally, skin prick tests performed on 28 P. judaica-sensitive patients showed that all of them recognized both isoforms and that allergenic epitopes present in Par j IA and IB are responsible for most of the allergenic activity of the whole extract.

Purification of Art v I, a relevant allergen of Artemisia vulgaris pollen.

An allergenic protein from Artemisia vulgaris pollen has been purified to homogeneity. Its molecular weight in native conditions is 47,000. The purified allergen, hereafter denominated Art v I, is a monomeric protein. It is a clinically relevant allergen since, at least, 70% of the individuals allergic to Artemisia vulgaris pollen have specific IgE in serum and in mast cells, demonstrated by ELISA and skin prick tests, respectively.

Synthesis and biological activity of photoactive derivatives of erythromycin.

Five photoactive derivatives of erythromycin have been synthesized by linking to 9(S)-aminoerythromycin either an aryl azide or a p-nitrophenyl ether. One derivative is an amide formed by reaction with (5-azido-2-formyl-phenoxy)acetic acid. Three derivatives are also amides, synthesized with 4-(p-nitroguaiacoxy)butanoic acid as a photoreactive group either directly or by interposing an amino acid (glycine or tyrosine). The last derivative is the product of the aldehyde condensation of aminoerythromycin with 10-(p-nitroguaiacoxy)decanal. Two of these derivatives can easily be made radioactive for affinity labeling studies either by reduction with [3H]borohydride (aryl azide derivative) or by 125I iodination (4-(p-nitroguaiacoxy)tyrosyl derivative). Although affected to different extents, the five erythromycin derivatives are biologically active and bind to the erythromycin-specific site on the bacterial ribosomes. In addition, the introduction of these groups changes the erythromycin inhibition pattern of peptide bond model reactions.

Identification of IgE-binding proteins from Lepidoglyphus destructor and production of monoclonal antibodies to a major allergen.

The allergen composition of one of the most important storage mites, Lepidoglyphus destructor, has been studied by immunodetection after SDS-PAGE with individual patient sera. An allergenic polypeptide of 14 kDa was identified with 95% of the sera. This major allergen was isolated in the supernatant of 60% ammonium sulfate salt precipitation of the whole extract, which was subsequently used to immunize BALB/c mice so as to produce monoclonal antibodies. Four mAbs recognizing molecules with IgE-binding ability were obtained. The specificity of the mAbs was assayed against different allergenic extracts, and the molecules recognized by them were characterized by immunoblotting. Two mAbs (Le5B5 and Le9E4) were directed to the 14-kDa allergen; the other two to several proteins of lesser allergenic significance.

Relationship between presence of Salmonella and indicators of faecal pollution in aquatic habitats.

The presence of Salmonella and its relationship with indicators of faecal pollution was investigated in aquatic habitats. The highest frequency was obtained in rivers (58.7% of samples) followed by freshwater reservoirs (14.8%) and sea water (5.9%). The sporadic presence of Salmonella (< 30%) on beaches with low concentrations of faecal streptococci (mean 25 CFU (100 ml)-1) may represent a potential risk for bathers in agreement with data found in previous epidemiological studies. Absence of Salmonella was observed only on beaches with very low densities (CFU (100 ml)-1) of indicator organisms (25 total coliforms, 13 faecal coliforms and 17 faecal streptococci).

Study of pancreatic secretion after subtotal gastrectomy followed by different methods of reconstruction.

To study the effect subtotal gastrectomies have on exocrine pancreatic function, 12 dogs were prepared with gastric and duodenal cannulas using a modified technique of Thomas. In our study protocol, we collected pancreatic juice by selective ductal cannulation after having applied two types of stimulation [tryptophan intraduodenally together with secretin intravenously (i.v.) and cholecystokinin i.v. together with secretin i.v.]. Our results did not show a significant increase in pancreatic juice after performing the gastrectomies. In both cases the concentration of bicarbonate decreased significantly in contrast to that in the control group. After Billroth I anastomosis, however, a significant decrease in the first periods of stimulation in relation to the gastrojejunostomy was appreciated. In the case of proteins, a loss of the dose/effect relation in these parameters was seen only after Billroth II anastomosis.

Cluster versus conventional immunotherapy in patients allergic to Dermatophagoides pteronyssinus: a controlled study of in vivo and in vitro parameters.

We designed a cluster schedule of immunotherapy for patients allergic to Dermatophagoides pteronyssinus which showed good safety and clinical efficacy. Here we compare the in vivo and in vitro changes with those of a conventional schedule in a controlled trial. Sixty-three patients were randomized as follows: 29 were treated with the cluster schedule, 15 with a conventional schedule and 19 received no immunotherapy. A standardized extract was used. Changes in in vivo parameters (skin prick test and conjunctival provocation test) and in in vitro parameters (IgE, IgG, IgG1 and IgG4 for the complete extract, Der p 1 and Der p 2) were measured before immunotherapy (T0), on reaching maintenance phase (T1), and after 6 (T2), 12 (T3) and 18 months of maintenance (T4). Cutaneous reactivity showed a significant decrease from T1 in both the cluster and conventional schedules, and conjunctival reactivity was also significantly lowered from T1 in these groups. Specific IgE decreased and specific IgG, IgG1 and IgG4 increased significantly from T1 in the cluster and conventional schedules. Neither of these parameters showed any changes in the group without immunotherapy. In conclusion, our cluster schedule induced changes in cutaneous and conjunctival reactivity and in immunological parameters that were similar to those achieved with the conventional schedule; these changes did not appear in patients who did not undergo immunotherapy.

Factors influencing the clinical picture and the differential sensitization to house dust mites and storage mites.

Sensitization to house dust mites and storage mites has been studied in a number of papers, but several environmental factors and clinical conditions that differently affect sensitization to these mites are still controversial. The purpose of this study was to determine the influence of climatic conditions, occupation and patient age in the differential sensitization to house dust and storage mites, and also to search for possible different symptoms caused by each group of mites. Eighty patients sensitized to mites but not to other inhalant allergens were studied by case history and by skin prick test and serum IgE to Dermatophagoides pteronyssinus. Dermatophagoides farinae, Lepidoglyphus destructor and Tyrophagus putrescentiae. Home conditions, including content of the allergens Der p 1, Der f 1, Lep d 2 and Tp, were determined for all patients. Human activities, such as farming or similar occupations, and humidity are conditions for preferential sensitization to storage mites, while we found no difference between living in rural or urban areas. Mean age for the onset of sensitization was 6.7 years for house dust mites and 18.7 years for storage mites. Conjunctivitis was more frequent in patients allergic to storage mites, whereas perioral syndrome (itching of the tongue and swelling of the lips) was only seen in patients sensitized to T. putrescentiae. We concluded that climatic and damp conditions and human activity, but not urban or rural living environments, influence the differential sensitization to house dust mites and storage mites.

Immunotherapy with a mass unit Parietaria judaica extract: a tolerance study with evidence of immunological changes to the major allergen Par j 1.

Specific immunotherapy with Parietaria judaica pollen extract has been proven to be effective in the treatment of patients with respiratory allergy induced by this pollen. Nevertheless, there is some controversy about its tolerability. We conducted an open uncontrolled study to evaluate the tolerability of an aluminium adsorbed P. judaica pollen extract whose major allergen (Par j 1) content was known. Changes in immunological parameters to a complete P. judaica extract and to a purified Par j 1 preparation were monitored. Twenty-one patients (12 women, 9 men; mean age 30.2 years) suffering from rhinoconjunctivitis and/or asthma due to Parietaria pollen were enrolled. The maximum dose was established at 10 BU/ml (0.6 microgram Par j 1). Skin and conjunctival reactivity as well as serum levels of specific IgE, IgG, IgG1 and IgG4 were evaluated before therapy (T0), when 1 BU was given (T1), 2 weeks after the maintenance dose was reached (T2) and after the pollen season (T3). Four-hundred and fifteen doses were administered during immunotherapy. Only one systemic reaction (0.24% of doses) and two local reactions were registered. Reactions occurred during the administration of the highest concentrated vial. Before immunotherapy, purified Par j 1 accounted for 94.2% of the cutaneous response elicited by the complete extract. A statistically significant decrease in cutaneous response was detected after 8 weeks of treatment. There were no significant changes in conjunctival reactivity throughout the study. Specific IgG, IgG1 and IgG4 showed a pronounced and significant increase during the study, while specific IgE levels initially decreased and increased after the pollen season. The kinetics of specific antibodies to P. judaica complete extract and purified Par j 1 showed a parallel trend. The present study demonstrates that immunotherapy with P. judaica extract is well tolerated in patients suffering from rhinoconjunctivitis and/or asthma due to Parietaria pollen. This therapy induces specific changes in the immunological response to P. judaica and to purified Par j 1. These changes can be detected at very early stages of therapy.