Proposal for encoding the surgical treatment in abdominal wall pathology based on a multidimensional analysis of history.

OBJECTIVE: To develop a multidimensional method that allows to identify different treatment concepts, techniques, protagonists, and their connections in surgical pathology of the abdominal wall throughout its historical development, serving as a basis or guide for the future. METHOD: First, an extensive and rigorous review of the literature was conducted to search for and group the different treatments described in the most common abdominal wall pathologies, including both groin and ventral hernias. Then, all treatment approaches were chronologically ordered and grouped according to their author, surgical approach, and method of approach. With all the information gathered, a table was created following a rational and multidimensional criterion that allows for the encoding of the set. RESULTS: 21 treatment modalities were identified and distributed into 8 groups. Additionally, 3 types of authors were detected: the creator, the innovator, and the popularizer. The assignment of values to different dimensions allowed us to obtain an alphanumeric code representative of the set. CONCLUSION: Multidimensional historical analysis allows analytical objectivity and set encoding. Its practical scope should be investigated.

Prevalence and clinical characteristics of symptomatic diffuse interstitial lung disease in rheumatoid arthritis in a Spanish population.

BACKGROUND AND OBJECTIVES: In Spain, epidemiological studies of the prevalence of diffuse interstitial lung disease (ILD) in rheumatoid arthritis (RA) are limited. Our objective was to estimate the prevalence of symptomatic ILD in RA and its characteristics in our area. MATERIALS AND METHODS: In our hospital's interdisciplinary rheumatology and pulmonology clinic, a prospective longitudinal observational study was designed in which we included RA with respiratory symptoms and ILD confirmed by high resolution computed tomography. RESULTS: Of the 2729 people with RA in our area, 47 had symptomatic ILD, estimating a prevalence of symptomatic ILD in RA of 1.72% (95% CI 1.26-2.29) with an age at diagnosis of RA of 57.3 ±â€¯13.3 years. It was more frequent in men, 60.6% had a history of smoking, and 84.3% and 84.7% had rheumatoid factor (RF) and anti-cyclic citrullinated peptide (Anti-CCP) antibodies, respectively. The most frequent pattern was usual interstitial pneumonitis (UIP), appearing in 28 (31.1%). Nonspecific interstitial pneumonia (NSIP) was more frequent in women, while the combined pulmonary fibrosis-emphysema (CPFE) syndrome presented exclusively in men. CONCLUSIONS: We have analysed the prevalence of symptomatic RA-ILD in our area, which is lower than expected, probably in relation to the definitions used. We have also described that the UIP pattern is the most frequent in RA in our environment, followed by the NSIP. Lastly, we have analysed the prevalence of CPFE in RA, which reaches 13%, for the first time.

Phosphoproteomic identification of a PDX-1/14-3-3ε interaction in pancreatic beta cells.

Glucose-dependent activation of the homeodomain transcription factor PDX-1 leads to its phosphorylation, to an increase in DNA binding capacity, and to NLS dependent translocation into the nucleus. To uncover unknown mediators of PDX-1 activation, PDX-1 interacting proteins were analysed by pull-down from (32)P-labelled, glucose-stimulated MIN6 cells. Recovered proteins were analysed by 2D gel electrophoresis and mass spectrometry. We identified 14-3-3ε as a novel PDX-1 binding protein and confirmed the interaction in vivo by Fluorescence Resonance Energy Transfer (FRET) analysis. We propose that 14-3-3ε interacts directly with PDX-1 to regulate its cellular distribution in pancreatic beta cells.

[Airway complications following lung transplantation - clinic, diagnosis, and interventional management]. / Atemwegskomplikationen nach Lungentransplantation - Klinik, Diagnose und interventionelle Behandlung.

Tracheobronchial complications following lung transplantation are defined as local structural or infectious alterations of the airways, which occur early or several months after lung transplantation (LTx). They preferentially develop in the region of the bronchial anastomosis. The most frequently reported complications are bronchial stenosis, bronchial dehiscence, exophytic excessive granulation tissue formation, tracheo-bronchomalacia, bronchial fistulas, and endobronchial infections. Airway complications are mainly attributed to ischaemia of the donor bronchus during the immediate post-transplant period. The most relevant risk factors for the development of airway complications include local infections, surgical techniques, and the immunosuppressive regimen. Thus, management of post-transplant bronchial complications requires early interventional bronchoscopic procedures including balloon bronchoplasty, cryotherapy, laser photoresection, endobronchial brachytherapy, and bronchial stents. In addition, antibiotic treatment, or non-invasive positive-pressure ventilation may be necessary. The procedures required depend on the time of occurrence, the type, and clinical relevance of the airway complication. This review summarises clinical presentation, risk factors, the diagnostic methods as well as management options for the most common LTx-associated airway complications.

Should we consider paranasal and chest computed tomography in severe asthma patients?

BACKGROUND: It is essential to recognize and treat findings that can simulate or worsen symptoms to improve asthma control and thereby to reduce costs. Guidelines highlight a paranasal (PS) and chest computed tomography (CT) scan as a tool for disease evaluation and, although they suggest its indication in patients whom presentation is atypical, there are not well-defined criteria. OBJECTIVES: To describe the most common findings in the PS and chest CT in severe asthma patients and to analyse the characteristics of asthmatics with the finding of nasal polyps or bronchiectasis. METHODS: We retrospectively reviewed the medical records of 161 adults with confirmed severe asthma who had undergone to PS and/or chest CT. Clinical data from their electronic health record and the findings from a PS and/or chest CT within the last five years were collected. RESULTS: In the PS CT, 70.5% of patients presented mucous thickening and 46.7% presented nasal polyps. Both findings were associated with male gender and level of blood eosinophils. In chest CT, 28% of individuals showed atelectasis, 16.5% air trapping, 17.7% affectation of the small airway, 11.6% pulmonary infiltrates and 10.4% emphysema. Bronchiectasis were identified in 60.4% of subjects, who were older and had poorer lung function. CONCLUSION: Paranasal and thoracic computed tomography are important tools in the treatment of severe asthma because they allow us to detect highly prevalent findings in this disease that can lead to poorer control of it.

Affinity capture mass spectrometry of biomarker proteins using peptide ligands from biopanning.

Affinity capture mass spectrometry was used to isolate and ionize protein A from Staphylococcus aureus from both a commercial source and cell culture lysate using matrix assisted laser desorption/ionization (MALDI) mass spectrometry. Two surfaces are compared: gold surfaces with immunoglobulin G covalently immobilized and silica surfaces with a covalently bound small peptide discovered via biopanning. A detection limit of 2.22 bacterial cells/mL of culture fluid was determined for the immobilized peptide surfaces. This study emphasizes the ability to use peptide ligands to effectively capture a biomarker protein out of a complex mixture. This demonstrates the potential to use biopanning to generate capture ligands for a large variety of target proteins and subsequently detect the captured protein using MALDI mass spectrometry.

[Interferon-alpha as treatment option in severe persistent uncontrolled bronchial asthma: an open label study]. / Interferon-alpha als Therapieoption bei der Behandlung des schwergradig persistierenden, unkontrollierten Asthma bronchiale: Eine offene Studie.

BACKGROUND: The purpose of this study was to evaluate the long-term safety and therapeutic effects of IFN-alpha in patients with severe persistent uncontrolled asthma on long-term oral glucocorticoid (GC) treatment. PATIENTS AND METHODS: The study included 16 patients (2 male, 14 female; age 39 years [range: 24 - 63]) with severe persistent asthma. Diagnosis and severity classification of asthma were established according to the guidelines of the "Deutsche Atemwegsliga". Eight patients stopped the therapy within 7 months due to side effects (n = 3), costs not covered by health insurance (n = 2), non-compliance (n = 2), and change of residence (n = 1). 8 patients (8 female, age 49 years [range: 35 - 68], duration of disease 16 years [range: 5 - 24]) were treated for at least 12 months with IFN-alpha (9 microg) 3 times/week. All patients were on oral glucocorticoids (GCs) for more than 5 years (average dose 17.5 [range: 5.0 - 64.0] mg/d). Clinical signs, lung function, need for reliever medication, number of emergency visits and hospitalisations and diary were assessed prior to and after 12 months of treatment. Data are given as percent of normal or median [range]. RESULTS: IFN-alpha improved lung function after 12 months: FEV1 64 vs. 75 %; FEV1/IVC 76 vs. 89 %; RV 153 % vs. 129 %; Rtot 193 vs. 111 % and morning PEF by 50 - 190 L/min. IFN-alpha also significantly reduced the use of reliever medication (10 [2 - 20] vs. 1 [0 - 3] puffs/d), nocturnal awakening (11 [4 - 30] vs. 1 [0 - 5]/month), emergency visits (7 [2 - 15] vs. 0 [0 - 5]/month) and hospitalisations (4 [1 - 8] vs. 0 [0 - 5]/year). In 5 patients the asthma attacks and nightly disturbances disappeared completely. The improvements were achieved despite a tapering of the oral GCs in all patients from 17.5 (5.0 - 64.0) to 2 (0 - 16) mg/d. In 5 patients GC treatment could be discontinued. The number of blood eosinophils decreased from 0.46 to 0.28 Gpt/L. Adverse events were transient and usually decreased within 3 to 4 weeks. Two patients developed an autoimmune thyreoiditis. CONCLUSION: In severe persistent, uncontrolled, and GC-dependent asthma, treatment with IFN-alpha leads to sustained clinical improvement and allows the reduction or discontinuation of oral GCs. Severe side effects may occur in isolated cases.

Inhibition of BCL11B expression leads to apoptosis of malignant but not normal mature T cells.

The B-cell chronic lymphocytic leukemia (CLL)/lymphoma 11B gene (BCL11B) encodes a Krüppel-like zinc-finger protein, which plays a crucial role in thymopoiesis and has been associated with hematopoietic malignancies. It was hypothesized that BCL11B may act as a tumor-suppressor gene, but its precise function has not yet been elucidated. Here, we demonstrate that the survival of human T-cell leukemia and lymphoma cell lines is critically dependent on Bcl11b. Suppression of Bcl11b by RNA interference selectively induced apoptosis in transformed T cells whereas normal mature T cells remained unaffected. The apoptosis was effected by simultaneous activation of death receptor-mediated and intrinsic apoptotic pathways, most likely as a result of tumor necrosis factor-related apoptosis-inducing ligand (TRAIL) upregulation and suppression of the Bcl-xL antiapoptotic protein. Our data indicate an antiapoptotic function of Bcl11b. The resistance of normal mature T lymphocytes to Bcl11b suppression-induced apoptosis and restricted expression pattern make it an attractive therapeutic target in T-cell malignancies.

Glucocorticoid receptor homodimers and glucocorticoid-mineralocorticoid receptor heterodimers form in the cytoplasm through alternative dimerization interfaces.

Steroid hormone receptors act to regulate specific gene transcription primarily as steroid-specific dimers bound to palindromic DNA response elements. DNA-dependent dimerization contacts mediated between the receptor DNA binding domains stabilize DNA binding. Additionally, some steroid receptors dimerize prior to their arrival on DNA through interactions mediated through the receptor ligand binding domain. In this report, we describe the steroid-induced homomeric interaction of the rat glucocorticoid receptor (GR) in solution in vivo. Our results demonstrate that GR interacts in solution at least as a dimer, and we have delimited this interaction to a novel interface within the hinge region of GR that appears to be both necessary and sufficient for direct binding. Strikingly, we also demonstrate an interaction between GR and the mineralocorticoid receptor in solution in vivo that is dependent on the ligand binding domain of GR alone and is separable from homodimerization of the glucocorticoid receptor. These results indicate that functional interactions between the glucocorticoid and mineralocorticoid receptors in activating specific gene transcription are probably more complex than has been previously appreciated.

[Establishing a protein signature from prostate tissue biopsies]. / Erstellung eines Proteinprofils aus Gewebeproben der Prostata.

The prostate-specific antigen test (PSA) has been a major factor contributing to a better management of prostate cancer. The low specificity limits its use in diagnosis especially in early detection of prostate cancer. Multiply expressed proteins need to be identified to establish a disease-specific protein signature that distinguishes between cancerous and noncancerous tissue. The first aim of our study is to identify differentially expressed proteins in both tissues using two-dimensional gel electrophoresis and subsequent mass spectrometry. We elucidated whether prostate biopsies are useful. First results have shown a different protein expression pattern in cancerous and noncancerous tissue. PCR revealed an increasing amount of mRNA for some upregulated proteins. We conclude that biopsies are useful material to establish protein expression patterns.

Reconstrucción poscirugía de Hartmann / Stoma reversal after the Hartmann's procedure

RESUMEN Antecedentes: la reconstrucciónn del tránsito intestinal luego de una operación de Hartmann es un procedimiento habitualmente complejo y con alta morbilidad. Objetivo: analizar la tasa de reconstrucción después de la cirugía de Hartmann y resultados posoperatorios en nuestra experiencia. Material y métodos: análisis retrospectivo de pacientes a los que se les practicó la reconstrucción del tránsito intestinal posterior a una cirugía de Hartmann en un período 16 años. Revisamos la bibliografía y nuestra base de datos. Luego traspasamos la información disponible a una grilla de datos construida con variables habitualmente analizadas en la literatura. Finalmente, analizamos los resultados mediante medidas básicas de tendencia central. Resultados: en 16 años realizamos 92 operaciones de Hartmann, de las cuales 69 (75%) llegaron a la reconstrucción. Edad promedio: 58 años. El 52% de los pacientes fueron hombres. La operación de Hartmann fue de urgencia en el 48% y 58% resultaron malignas. Tiempo transcurrido hasta la reconstrucción: en promedio, 9 meses, y el 90% (N 62) de los casos se realizó por vía laparoscópica. Morbilidad general 38% y ajustada a los grados III y IV de Clavien-Dindo fue 11,5%. No hubo mortalidad. Conclusión: los resultados obtenidos son semejantes a los publicados y nuestra experiencia nos motiva a continuar eligiendo el abordaje laparoscópico.

ABSTRACT Background: Background: Stoma reversal after Hartman's operation is usually a complex procedure and is associated high morbidity. Objective: To analyze the rate of reversal after the Hartmann's procedure and the postoperative outcomes in our experience. Material and methods: We conducted a retrospective analysis of patients undergoing reversal after the Hartmann's procedure over a 16-year period with review of the literature and of our database and transferred the available information to a data grid constructed with variables commonly analyzed in the literature. Finally, we analyzed the results using basic measures of central tendency. Results: Over a 16-year period, we performed 92 Hartmann's operations; 69 (75%) reached the reversal stage. Mean age was 58 years and 52% were men. Forty-eight percent of the Hartmann's procedures were emergency surgeries and 58% were due to cancer. Mean time to reversal was 9 months and 90% (n = 62) were laparoscopic procedures. Overall morbidity and adjusted for complications grade III and IV of the Clavien-Dindo classification were 38% and 11.5%, respectively. None of the patients died. Conclusion: The results obtained are similar to those published and our experience motivates us to continue choosing the laparoscopic approach.

Tumor necrosis factor alpha induces the expression of the nuclear protein p8 via a novel NF kappaB binding site within the promoter.

p8 is a widely expressed HMG-I/Y-like transcription factor which is involved in regulating cell proliferation and tissue stress. Several studies describe a strong upregulation of p8 expression during inflammatory processes like pancreatitis and LPS-induced sepsis. Here we demonstrate that TNFalpha, which is an important inducer of innate defence against gram-negative bacteria, significantly stimulates p8 protein production in H4IIE rat hepatoma cells within 2 hours. Since a putative NF kappaB motif has been described, we further tested whether TNFalpha stimulates p8 expression via activation of NF kappaB. We characterized the TNFalpha-induced binding of NF kappaB to this motif. We show that the TNFalpha-induced NF kappaB pathway contributes to the induction of p8 during pancreatitis and LPS-induced inflammation.

Basal level glucose-6-phosphatase gene transcription requires binding sites for Sp family proteins within the gene promoter.

The significance of two regions (SpA: -19 to -11 and SpB: -63 to -55) within the human glucose-6-phosphatase (G6Pase) gene promoter for gene expression was examined. The mutation of SpA and SpB together, but not alone, decreased G6Pase promoter activity. Electromobility shift assays showed that SpA and SpB were able to bind the transcription factors Sp1 and Sp3.

Quantitative real-time polymerase chain reaction for malaria diagnosis and its use in malaria vaccine clinical trials.

The demand for an effective malaria vaccine is high, with millions of people being affected by the disease every year. A large variety of potential vaccines are under investigation worldwide, and when tested in clinical trials, researchers need to extract as much data as possible from every vaccinated and control volunteer. The use of quantitative real-time polymerase chain reaction (PCR), carried out in real-time during the clinical trials of vaccines designed to act against the liver stage of the parasite's life cycle, provides more information than the gold standard method of microscopy alone and increases both safety and accuracy. PCR can detect malaria parasites in the blood up to 5 days before experienced microscopists see parasites on blood films, with a sensitivity of 20 parasites/mL blood. This PCR method has so far been used to follow 137 vaccinee and control volunteers in Phase IIa trials in Oxford and on 220 volunteer samples during a Phase IIb field trial in The Gambia.

The Tet-On system in transgenic mice: inhibition of the mouse pdx-1 gene activity by antisense RNA expression in pancreatic beta-cells.

To elucidate the function of pancreas duodenal homeobox 1 (PDX-1; insulin promoter factor 1/somatostatin transcription factor 1/islet duodenum homeobox 1/insulin upstream factor 1) in differentiated beta-cells of adult animals we generated transgenic mice using the Tet-On system. Inducible expression of an antisense RNA should down-regulate the PDX-1 protein. The selective and continuous inhibition of PDX-1 gene expression should impair the expression of PDX-1 dependent beta-cell specific genes. A gene switch such as the Tet-On system provides a powerful tool to analyze eukaryotic gene expression and function in transgenic mice. The original Tet system contained two transcriptional units, transactivator and target of control, on two plasmids. We combined the two transcriptional units on a single DNA molecule. The transactivator was placed under control of the mouse insulin promoter. The tet responsive element, driving the gene of interest, was inserted further down-stream into the same vector. The tet regulatory system in this approach permitted a tissue-specific and a doxycycline-inducible control of PDX-1 expression in transgenic mice. The expression of glucose transporter 2 and glucokinase was markedly reduced in dox-treated transgenic mice. In contrast, the number of insulin- and amylin-expressing cells was only slightly decreased, whereas the expression of glucagon was increased distinctly in islets of these mice. Furthermore, the exposure to doxycycline resulted in a progressive impairment of glucose tolerance. The characterization of our transgenic mouse model demonstrates the suitability of the Tet-On system for analyzing physiological consequences emerging from a stepwise decrease in a given protein. Using this system we confirmed the essential role of PDX-1 in pancreatic islets and demonstrated that an antisense-mediated PDX-1 deficiency provokes a beta-cell dysfunction.

Effect of dietary fats and carbohydrate on blood pressure of mildly hypertensive patients.

The effect on blood pressure (BP) of replacing dietary saturated fat with either polyunsaturated fat (linoleic acid) or carbohydrate was studied in 21 untreated mildly hypertensive patients. In a randomized, double-blind, crossover protocol, all subjects received dietary supplements of cream, safflower oil, and carbohydrate in random sequence, each prepared in flavored yogurt or milk. Each supplement was administered for 6 weeks and followed by a 4-week washout period of no supplementation. Dietary linoleic acid increased from 4.6 to 13% of energy intake when the safflower oil replaced cream, while saturated fat decreased from 16 to 10%. Total fat intake was 37 to 38% during the cream and safflower oil periods but was 28% during the carbohydrate period. Compliance with the diets was demonstrated by significant changes in fasting plasma fatty acid measurements. Mean clinic BP was 135 +/- 9/93 +/- 6 mm Hg at baseline. There were no significant differences in BP measured in the clinic or at home among the three dietary periods. The protocol had more than 80% power to detect a mean effect of diet of 3 mm Hg systolic or 2 mm Hg diastolic BP. Therefore, replacing dietary saturated fat with carbohydrate or with linoleic acid does not affect BP in subjects with mild hypertension.

Positive and negative regulatory elements are involved in transcriptional control of the rat glucokinase gene in the insulin producing cell line HIT M2.2.2.

Nested deletion mutants of the 5' flanking region of the beta-cell transcription unit of the rat glucokinase gene (r beta GK) were fused to the CAT-reporter gene. Transient expression studies in HIT M2.2.2 and BHK21 cells revealed a distal (upstream of -359) and a proximal promoter region (between -278/-49) harbouring positive and negative regulatory elements. DNaseI footprinting revealed three protected areas between nucleotides -190 and -60. DNA-elements playing a crucial role in transcriptional control of the insulin genes (IEB- and CT-motifs) have been detected within the proximal promoter region and contribute to beta-cell specific gene regulation. 3' deletion analysis revealed that DNA-elements located downstream from transcription initiation sites (up to +123) contribute to transcriptional regulation.

CT-boxes are involved in control of the rat insulin II gene expression.

Expression of the rat insulin II gene is controlled mainly at the level of transcription initiation by multiple factors binding to specific cis-acting DNA-elements in the regulatory region. We have shown that two elements (CT-motifs) located between nucleotides -83 and -76 (CT-1) and -204 and -197 (CT-2) are involved in transcriptional regulation in the insulin-producing cell line HIT M2.2.2. Transient expression analysis of 5'-deletion as well as block replacement mutants revealed that CT-1 and CT-2 are mutational sensitive. Gel mobility shift assays showed that both motifs bind similar nuclear factors. Our results suggest the involvement of a third CT-motif located directly upstream of CT-2 on the complementary strand.

Functional analysis of a newly identified TAAT-box of the rat insulin-II gene promoter.

Transcriptional regulation of insulin gene expression is achieved by an interplay of tissue-specific and ubiquitous cis- and trans-acting elements. E-box like motifs and TAAT-motifs were shown to play a crucial role in initiating insulin gene transcription. Studying the AT-rich region of the rat insulin-II promoter between nucleotides -212 and -196, we observed a base difference at -211, an adenosine instead of a cytidine, compared to the previously reported sequence (EMBL Accession No. J00748). Sequence analysis of promoter fragments from different rat strains showed that adenosine at position -211 represents the wild type (EMBL Accession No. X82162). This base exchange leads to the formation of an additional TAAT-motif, i.e. TAAT3, at the complementary DNA strand directly upstream of the previously studied TAAT2 motif, formerly named CT-2. Here we show that the newly identified motif TAAT3 is involved in (i) transcriptional control in vivo, (ii) in vitro DNA/protein interactions, and that (iii) TAAT1, TAAT2 and TAAT3 are binding sites for the homeodomain-containing factor IPF-1.