Transrectal high-intensity focused ultrasound (HIFU) for management of rectosigmoid deep infiltrating endometriosis: results of Phase-I clinical trial.

OBJECTIVES: Deep infiltrating endometriosis (DIE) of the rectosigmoid is associated with painful symptoms. When medical treatment is ineffective, surgical resection remains the standard treatment, despite significant risk of adverse events. High-intensity focused ultrasound (HIFU) is a minimally invasive ablative procedure. Focal One® is a transrectal HIFU (TR-HIFU) device used in prostate cancer treatment. The primary objective of this study was to confirm the feasibility of treatment with TR-HIFU in patients presenting with posterior DIE with rectosigmoid involvement. We also assessed its safety and clinical efficacy in this context. METHODS: This was a non-controlled, prospective, Phase-I clinical trial in a French University Hospital which is a multidisciplinary center for management of endometriosis. Included were patients older than 25 years, without plans to conceive within 6 months, who presented with a single lesion of posterior DIE, with rectosigmoid invasion, after failure of hormonal therapy. All lesions were assessed preoperatively using transvaginal sonography and magnetic resonance imaging. Patients completed questionnaires on gynecological and intestinal symptoms (similar to a visual analog scale (VAS)), and on quality of life (Medical Outcomes Study 36-item short-form survey (SF-36) and, for the second half of patients recruited, symptom scoring system for constipation (KESS), female sexual function index (FSFI) and endometriosis health profile short-version score (EHP-5)), before, and at 1, 3 and 6 months after, TR-HIFU treatment with a Focal One real-time ultrasound-guided HIFU device. RESULTS: Twenty-three consecutive patients were included in the study between September 2015 and October 2019. All 23 lesions were visualized, giving a detection rate of 100%. Twenty lesions were treated ('feasibility rate', 87.0%): in 13 the whole lesion was treated and in seven the lesion was treated partially. The mean duration of the TR-HIFU procedure was 55.6 min. We observed a significant improvement in VAS score at 6 months, with differences relative to preoperative scores as follows, for: dysmenorrhea (-3.6, P = 0.004), dyspareunia (-2.4, P = 0.006), diarrhea (-3.0, P = 0.006), constipation (-3.0, P = 0.002), dyschezia (-3.2, P = 0.003), false urge to defecate (-3.3, P = 0.007), posterior pelvic pain (-3.8, P = 0.002) and asthenia (-3.8, P = 0.002). There was also a significant improvement in the SF-36 score, with an increase at 6 months relative to the preoperative score in both the physical component summary (+ 9.3%, P = 0.002) and mental component summary (+ 10.9%, P = 0.017). No major complications occurred during or after any procedure. CONCLUSIONS: TR-HIFU therapy for posterior DIE is feasible. If its efficacy and safety are confirmed, it could be a minimally invasive alternative to surgery for the treatment of rectosigmoid endometriosis. © 2019 Authors. Ultrasound in Obstetrics & Gynecology published by John Wiley & Sons Ltd on behalf of International Society of Ultrasound in Obstetrics and Gynecology.

Exposure to dogs but not cats is associated to a decrease in the prevalence in atopic dermatitis amongst school-children.

INTRODUCTION: The association regarding the exposure to pets, especially cats and dogs, and the prevalence of allergic diseases is inconsistent. OBJECTIVE: We analyzed the role played by early exposure to dogs or cats in the prevalence of allergic diseases amongst school-aged children. METHOD: Through a cross-sectional study, we examined 756 children, aged 6-7; these candidates were selected through cluster sampling. We inquired about the exposure that these children had had to dogs and cats, and whether these pets spent most of their time indoors or outdoors during the first year of the child's life. In order to identify the prevalence of allergic diseases and their symptoms, each child's parent completed the International Study of Asthma and Allergies in Childhood questionnaire. RESULTS: Exposure to outdoor dogs was associated to nocturnal coughing, odds ratio (OR) 0.64, with a confidence interval of 95% (95% CI) 0.43-0.95 and with atopic dermatitis (OR: 0.39; 95% CI: 0.20-0.76). Interestingly, exposure to outdoor cats was associated to nocturnal coughing (OR: 0.51; 95% CI: 0.32-0.83) and current rhinitis symptoms (OR: 0.59; 95% CI 0.36-0.97). After carrying out the multivariate analyses, only exposure to dogs, both indoor and outdoor, was significantly associated to a decrease in the prevalence of atopic dermatitis OR 0.40 (95% CI: 0.20-0.79) and OR 0.38 (95% CI: 0.18-0.83), respectively. CONCLUSION: Our findings suggest that exposure to dogs, whether they be indoor or outdoor pets, is associated to a decreased prevalence in atopic dermatitis.

From vasospasm to early brain injury: new frontiers in subarachnoid haemorrhage research.

INTRODUCTION: Delayed vasospasm has traditionally been considered the most important determinant of poor outcome after subarachnoid haemorrhage (SAH). Consequently, most of the research and therapies are directed towards reducing the incidence of vasospasm (VSP). To date, however, clinical trials based on this strategy have not delivered a definitive treatment for preventing or reducing brain injury after SAH. This fact has caused a paradigm shift in research, which now focuses on early brain injury (EBI) occurring in the first 72 hours after SAH. It has also changed the idea of VSP's role in brain damage, and suggests the need for re-evaluating the pathophysiological process of SAH. DEVELOPMENT: This review examines the current state of knowledge on the pathophysiological mechanisms associated with EBI and summarises the diagnostic options currently available. CONCLUSION: It seems that the research approach needs to be changed so that investigators will focus on prevention of EBI, reduction of secondary brain complications and ultimately, the optimisation neurological outcome.

A functional study of nucleocytoplasmic transport signals of the EhNCABP166 protein from Entamoeba histolytica.

EhNCABP166 is an Entamoeba histolytica actin-binding protein that localizes to the nucleus and cytoplasm. Bioinformatic analysis of the EhNCABP166 amino acid sequence shows the presence of 3 bipartite nuclear localization signals (NLS) and a nuclear export signal (NES). The present study aimed to investigate the functionality of these signals in 3 ways. First, we fused each potential NLS to a cytoplasmic domain of ehFLN to determine whether the localization of this domain could be altered by the presence of the NLSs. Furthermore, the localization of each domain of EhNCABP166 was determined. Similarly, we generated mutations in the first block of bipartite signals from the domains that contained these signals. Additionally, we added an NES to 2 constructs that were then evaluated. We confirmed the intranuclear localization of EhNCABP166 using transmission electron microscopy. Fusion of each NLS resulted in shuttling of the cytoplasmic domain to the nucleus. With the exception of 2 domains, all of the evaluated domains localized within the nucleus. A mutation in the first block of bipartite signals affected the localization of the domains containing an NLS. The addition of an NES shifted the localization of these domains to the cytoplasm. The results presented here establish EhNCABP166 as a protein containing functional nuclear localization signals and a nuclear export signal.

Role of Pegylated Interferon in Patients with Chronic Liver Diseases in the Context of SARS-CoV-2 Infection.

The coronavirus 2019 (COVID-19) pandemic has resulted in 168 million cases and about 3.5 million deaths (as of May 26, 2021) during the last 18 months. These 18 months of the COVID-19 pandemic have been characterized by phases or waves of new cases, the emergence of new variants of the deadly virus, and several new complications. After providing emergency approval to several drugs and adherence to several public health measures with frequent full and partial lockdowns, the incidence of new severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) could not be contained till now on a global basis. Although prophylactic vaccines have inspired optimism, the scarcity of vaccines and several vaccine-related regulations indicate that the vaccine's benefit would not be reaching the people of developing countries anytime soon. In the course of our clinical practice, we used pegylated interferon (Peg-IFN) in 35 patients with chronic liver diseases (CLD), and we found that only two of them were infected with SARS-CoV-2 that was mild in nature. These two patients with CLD have a mild course of disease cured without any specific therapy. Patients with CLD are usually immune-compromised. However, three CLD patients remained free of SARS-CoV-2 although they had COVID-19 patients among their family members. Next, we accomplished two studies for assessing the immune-modulatory capacities of Peg-IFN, 1 and 12 injections following administration of Peg-IFN. The data revealed that peripheral blood mononuclear cells (PBMCs) of Peg-IFN-administered CLD patients produced significantly higher levels of some cytokines of innate immunity in comparison with the cytokines produced by PBMC of CLD patients before Peg-IFN intake. The pattern of cytokine responses and absence of infection of SARS-CoV-2 in 33 of 35 CLD patients represent some preliminary observations indicating a possible role of Peg-IFN in patients with CLD. The study may be extended to other chronic infections and cancers in which patients receive Peg-IFN. The role of Peg-IFN for pre- or postexposure prophylaxis in the acquisition of SARS-CoV-2 infection and influencing the natural course of COVID-19 remains to be clarified. HOW TO CITE THIS ARTICLE: Akbar SMF, Mahtab MA, Aguilar JC, et al. Role of Pegylated Interferon in Patients with Chronic Liver Diseases in the Context of SARS-CoV-2 Infection. Euroasian J Hepato-Gastroenterol 2021;11(1):27-31.

Protection against invasive amebiasis by a single monoclonal antibody directed against a lipophosphoglycan antigen localized on the surface of Entamoeba histolytica.

A panel of monoclonal antibodies was raised from mice immunized with a membrane preparation from Entamoeba histolytica, the pathogenic species causing invasive amebiasis in humans. Antibody EH5 gave a polydisperse band in immunoblots from membrane preparations from different E. histolytica strains, and a much weaker signal from two strains of the nonpathogenic species Entamoeba dispar. Although the exact chemical structure of the EH5 antigen is not yet known, the ability of the antigen to be metabolically radiolabeled with [32P]phosphate or [3H]glucose, its sensitivity to digestion by mild acid and phosphatidylinositol-specific phospholipase C, and its specific extraction from E. histolytica trophozoites by a method used to prepare lipophosphoglycans from Leishmania showed that it could be classified as an amebal lipophosphoglycan. Confocal immunofluorescence and immunogold labeling of trophozoites localized the antigen on the outer face of the plasma membrane and on the inner face of internal vesicle membranes. Antibody EH5 strongly agglutinated amebas in a similar way to concanavalin A (Con A), and Con A bound to immunoaffinity-purified EH5 antigen. Therefore, surface lipophosphoglycans may play an important role in the preferential agglutination of pathogenic amebas by Con A. The protective ability of antibody EH5 was tested in a passive immunization experiment in a severe combined immunodeficient (SCID) mouse model. Intrahepatic challenge of animals after administration of an isotype-matched control antibody or without treatment led to the development of a liver abscess in all cases, whereas 11 out of 12 animals immunized with the EH5 antibody developed no liver abscess. Our results demonstrate the importance and, for the first time, the protective capacity of glycan antigens on the surface of the amebas.

Structure and content of the Entamoeba histolytica genome.

The intestinal parasite Entamoeba histolytica is one of the first protists for which a draft genome sequence has been published. Although the genome is still incomplete, it is unlikely that many genes are missing from the list of those already identified. In this chapter we summarise the features of the genome as they are currently understood and provide previously unpublished analyses of many of the genes.

Signaling through the phosphatidylinositol 3-kinase regulates mechanotaxis induced by local low magnetic forces in Entamoeba histolytica.

In micro-organisms, as well as in metazoan cells, cellular polarization and directed migration are finely regulated by external stimuli, including mechanical stresses. The mechanisms sustaining the transduction of such external stresses into intracellular biochemical signals remain mainly unknown. Using an external magnetic tip, we generated a magnetic field gradient that allows migration analysis of cells submitted to local low-intensity magnetic forces (50 pN). We applied our system to the amoeba Entamoeba histolytica. Indeed, motility and chemotaxis are key activities that allow this parasite to invade and destroy the human tissues during amoebiasis. The magnetic force was applied either inside the cytoplasm or externally at the rear pole of the amoeba. We observed that the application of an intracellular force did not affect cell polarization and migration, whereas the application of the force at the rear pole of the cell induced a persistent polarization and strongly directional motion, almost directly opposed to the magnetic force. This phenomenon was completely abolished when phosphatidylinositol 3-kinase activity was inhibited by wortmanin. This result demonstrated that the applied mechanical stimulus was transduced and amplified into an intracellular biochemical signal, a process that allows such low-intensity force to strongly modify the migration behavior of the cell.

Virulence and functions of myosin II are inhibited by overexpression of light meromyosin in Entamoeba histolytica.

Several changes in cell morphology take place during the capping of surface receptors in Entamoeba histolytica. The amoebae develop the uroid, an appendage formed by membrane invaginations, which accumulates ligand-receptor complexes resulting from the capping process. Membrane shedding is particularly active in the uroid region and leads to the elimination of accumulated ligands. This appendage has been postulated to participate in parasitic defense mechanisms against the host immune response, because it eliminates complement and specific antibodies bound to the amoeba surface. The involvement of myosin II in the capping process of surface receptors has been suggested by experiments showing that drugs that affect myosin II heavy-chain phosphorylation prevent this activity. To understand the role of this mechanoenzyme in surface receptor capping, a myosin II dominant negative strain was constructed. This mutant is the first genetically engineered cytoskeleton-deficient strain of E. histolytica. It was obtained by overexpressing the light meromyosin domain, which is essential for myosin II filament formation. E. histolytica overexpressing light meromyosin domain displayed a myosin II null phenotype characterized by abnormal movement, failure to form the uroid, and failure to undergo the capping process after treatment with concanavalin A. In addition, the amoebic cytotoxic capacities of the transfectants on human colon cells was dramatically reduced, indicating a role for cytoskeleton in parasite pathogenicity.

Role of signalling and cytoskeletal rearrangements in the pathogenesis of Entamoeba histolytica.

Virulence of Entamoeba histolytica is characterized by intestinal tissue invasion, engulfment of host cells and the formation of liver abscesses. The actin-rich cytoskeleton of the amoeba allows rapid changes in morphology in response to signals from external stimuli. Cellular and molecular studies have described some of the proteins that participate in signalling and in cytoskeletal changes.

In vitro transcription from Bacillus subtilis nucleoids by homologous RNA polymerase.

The effect of DNA structural features on RNA synthesis was investigated. Purified Bacillus subtilis nucleoids templated from vegetative cells were transcribed by the homologous RNA polymerase using Hg-UTP as one of the nucleotide substrates. Low RNA polymerase/DNA ratios were used during transcription in order to avoid nonspecific initiations. The rate of synthesis of total RNA was 40% greater on nucleoid templates than on naked DNA. The proportion of asymmetric transcript synthesized on nucleoid templates (HvsL strand transcripts) was close to that observed in vivo, whereas with naked DNA this value was at least 3-times lower. The percentage of rRNA relative to the total RNA, synthesized in the in vitro system with the nucleoid template, approaches the rate of the in vivo transcription. The size of the RNA synthesized on nucleoids was large and heterogeneous while with naked DNA it was homogeneous and of about 6 S. Our results suggest that the supercoiled, folded nucleoids may retain some of the structural features responsible for the regulation of RNA synthesis in vivo.

Genetic analysis of fusion recombinants in Bacillus subtilis: function of the recE gene.

Bacillus subtilis protoplast fusion allows the study of the genetic recombination of an entire procaryotic genome. Protoplasts from bacterial strains marked genetically by chromosomal mutations were fused using polyethylene glycol and the regenerated cells analyzed. Recombinants represent 19.3% of heterozygotic cells; they are haploids. Individual characterization of clones show a unique particular phenotype in each colony suggesting that recombination takes place immediately after fusion, probably before the first cellular division. Recombination occurs in the whole chromosome; in one-third of the cases both reciprocal recombinants could be shown in the colony. The genetic interval that includes the chromosome replication origin shows the highest recombination level. Our results suggest that the RecE protein accounts for most of the fused protoplast recombination; however, some "replication origin-specific" recombination events were independent of the recE gene product.

Cell polarization and adhesion in a motile pathogenic protozoan: role and fate of the Entamoeba histolytica Gal/GalNAc lectin.

The human pathogenic protozoan Entamoeba histolytica is a motile cell polarized into a front pseudopod and a rear uroid. The amoebic Gal/GalNAc surface lectin is a major adhesion molecule composed of an immunodominant 170-kDa heavy subunit, mostly extracellular except for a short cytoplasmic tail, and of an extracellular light subunit. The binding of multivalent ligands triggers lectin capping and recruitment to the uroid. The properties of the Gal/GalNAc lectin and its role in amoeba adhesion and uroid polarization are reviewed in the context of the molecular mechanisms underlying cell polarization and locomotion.

Identification and characterization of a myosin heavy chain gene (mhcA) from the human parasitic pathogen Entamoeba histolytica.

The mhcA gene from the human pathogen Entamoeba histolytica was identified using the polymerase chain reaction. It is a single copy gene expressed as a 6.4-kb mRNA. The deduced MhcA protein sequence is highly similar to myosin II from both Dictyostelium discoideum and Acanthamoeba castellanii. The globular head domain of MhcA contains the specific regions involved in ATP binding, actin binding, and interaction with myosin light chain. The tail domain is organized in an alpha-helical coiled coil structure, which suggests that MhcA is an alpha-fibrous protein. The coiled coil is interrupted by two prolines indicating that like other myosins, either from smooth muscle or from non-muscle cells, the tail of MhcA folds twice on itself. In addition, MhcA presents sequence similarities with the heavy chain phosphorylation sites of smooth and non-muscle vertebrate myosins.

Molecular characterization of myosin IB from the lower eukaryote Entamoeba histolytica, a human parasite.

The complete amino acid sequence of the Entamoeba histolytica unconventional myosin IB (Eh-myosin IB) is reported. Sequencing of overlapping cDNA fragments reveals a single open reading frame which predicts a 130 kDa protein of 1049 aa. Eh-myosin IB presents the three characteristics domains of myosins I subclass 1. This protein presents homology with myosins IB from other amoebae, but striking homologies with vertebrate unconventional myosins were also observed. The predicted actin and ATP-binding sites are located in the head domain. The heavy chain phosphorylation region is homologous to metazoan myosins I with an acidic residue present at the phosphorylation site. In the neck domain, an IQ motif indicates potential binding of calmodulin to the myosin I heavy chain. In the tail of Eh-myosin IB the three characteristic regions of myosin I are found. A putative membrane binding domain a very short domain rich in alanine and proline we demonstrate to be functional for actin binding, and the src-homology 3 domain. The Entamoeba histolytica myosin IB is the first unconventional myosin so far described in a lower eukaryote.

Identification of a myosin heavy chain gene from Entamoeba histolytica.

The myosin heavy chain gene A, mhcA, from E. histolytica was identified by polymerase chain reaction. There is only one copy of this gene on the ameba genome. The mhcA gene encodes a 6 kb messenger RNA. The predicted protein sequence is homologous to myosin II from both, Dictyostelium discoideum and Acanthamoeba castellanii. Specific areas of the N-terminal region of myosin II are highly conserved reflecting a similarity in functional domains, like the ATP or the actin binding sites.

[Food intake, dietary habits and nutritional status of the population of Reus (X): Evolution of the diet and the contribution of macronutrients to energy intake (1983-1999) by age and sex]. / Consumo, hábitos alimentarios y estado nutricional de la población de Reus (X): Evolución de la ingestión alimentaria y de la contribución de los macronutrientes al aporte energético (1983-1999), según edad y sexo.

BACKGROUND: To study the evolution of the diet and the nutritional intake between 1983 and 1999, by age and sex. SUBJECTS AND METHODS: We performed a series of analysis of the food intake on a representative sample of the population of Reus (aged 10-69 years). Dietary intake was estimated using the 24-hours recall method. In 1999 the sample size was 839 individuals, 41% of them having taken part in the studies since 1983. Results are shown as mean (standard deviation). RESULTS: In 1999, the energy intake was 2524 (582) kcal in men aged 35-44 years (n = 57), and 1827 (490) kcal in women (n = 95) (p < 0.001 between sexes). The energy intake decreases with age (significant trend [p < 0.001] between 15-69 years) and it is higher in men than in women in all the age groups studied. When comparing similar age groups, we observe that this energy intake has not changed significantly since 1983. Between 1983 and 1999 the contribution of the macronutrients to the total energy intake has become more similar between ages and sexes. In 1999, in men aged 35-44 years a 15.6% of the energy came from proteins, 42% from fat and 42.5% from carbohydrates; in women of the same age: a 17.3% of the energy came from proteins, 42.4% from fat and 40.3% from carbohydrates). During this period we observed, however, remarkable changes in the diet which imply the increasing participation of the dairy products, vegetables and meat in the energy intake, or the significant decrease of the role of tubers, eggs and visible fat. CONCLUSIONS: Our population maintains an energy intake without significant changes, and it presents a trend towards a progressive uniformity of the nutritional balance between the different ages and sexes, although there are significant changes in some components of the diet.

[Assessment of the acceptance of set meals served at the "Sant Joan de Reus" University Hospital]. / Evaluación de la aceptación de los menús servidos en el Hospital Universitari de Sant Joan de Reus.

Despite the fact that, in most of the series published, cancer is the most frequent base pathology for the indication of home parenteral nutrition (HPN), the use of this technique in terminally-ill patients is still a controversial issue. Our goal has been to review the evolution of cancer patients with HPN treatment from "La Paz" Hospital with a view to studying the indication, evolution and complications. We review a total of 9 terminal oncological patients who had been treated with HPN between January 2000 and December 2002, with a mean age of 60.4 (44-81) years, the most common base cancer was gastric adenocarcinoma (44%). Intestinal obstruction in the context of peritoneal carcinomatosis was the reason for indicating HPN in 89% of cases and the median survival time was 71 (23-131) days. Catheter infection was the most frequent complication with 1.4 episodes/patient. The existence of a Home Support Team meant follow-up of patients was easier, with HPN being estimated as the treatment provided in 67% of cases. 56% of the patients were not sufficiently informed as to their underlying illness. Although HPN is one more therapeutic resource, which may or may not be used in some terminal oncological patients, we must refine the indication as much as possible to take into account a series of "systematic guarantees" including fulfilment of pertinent clinical criteria, informed consent and the adoption of a collective decision with the involvement of all the professionals monitoring the patient. We propose an action algorithm to help in improving the decision-taking process in these patients.

A karyological study of the spirurid nematode Mastophorus muris (Nematoda: Spirocercidae).

The karyotype of Mastophorus muris (Gmelin, 1790) comprises four pairs of small autosomal chromosomes and two larger sex X chromosomes in females or one X chromosome in males (2n = 8 + XX/XO). All pairs of chromosomes possess rather uniform morphology without distinct primary or secondary constrictions. No heterochromatin bands were found by C-banding analysis. The absolute chromosome length ranges from 4.02 to 2.24 microns. The mean total length of the haploid complement is 14.34 microns. The course of gametogenesis represents a typical pattern common in the order Spirurida. The recently available karyotypes of spirurid nematodes have been reviewed.