Spatial control of gene expression in flies using bacterially derived binary transactivation systems.

Controlling gene expression is an instrumental tool for biotechnology, as it enables the dissection of gene function, affording precise spatial-temporal resolution. To generate this control, binary transactivational systems have been used employing a modular activator consisting of a DNA binding domain(s) fused to activation domain(s). For fly genetics, many binary transactivational systems have been exploited in vivo; however, as the study of complex problems often requires multiple systems that can be used in parallel, there is a need to identify additional bipartite genetic systems. To expand this molecular genetic toolbox, we tested multiple bacterially derived binary transactivational systems in Drosophila melanogaster including the p-CymR operon from Pseudomonas putida, PipR operon from Streptomyces coelicolor, TtgR operon from Pseudomonas putida and the VanR operon from Caulobacter crescentus. Our work provides the first characterization of these systems in an animal model in vivo. For each system, we demonstrate robust tissue-specific spatial transactivation of reporter gene expression, enabling future studies to exploit these transactivational systems for molecular genetic studies.

Effect of homeostatic T-cell proliferation in the vaccine responsiveness against influenza in elderly people.

BACKGROUND: Seasonal influenza virus infection is a significant cause of morbimortality in the elderly. However, there is poor vaccine efficacy in this population due to immunosenescence. We aimed to explore several homeostatic parameters in the elderly that could impact influenza vaccine responsiveness. METHODS: Subjects (> 60 years old) who were vaccinated against influenza virus were included, and the vaccine response was measured by a haemagglutination inhibition (HAI) test. At baseline, peripheral CD4 and CD8 T-cells were phenotypically characterized. Thymic function and the levels of different inflammation-related biomarkers, including Lipopolysaccharide Binding Protein (LBP) and anti-cytomegalovirus (CMV) IgG antibodies, were also measured. RESULTS: Influenza vaccine non-responders showed a tendency of higher frequency of regulatory T-cells (Tregs) before vaccination than responders (1.49 [1.08-1.85] vs. 1.12 [0.94-1.63], respectively, p = 0.061), as well as higher expression of the proliferation marker Ki67 in Tregs and different CD4 and CD8 T-cell maturational subsets. The levels of inflammation-related biomarkers correlated with the frequencies of different proliferating T-cell subsets and with thymic function (e.g., thymic function with D-dimers, r = - 0.442, p = 0.001). CONCLUSIONS: Age-related homeostatic dysregulation involving the proliferation of CD4 and CD8 T-cell subsets, including Tregs, was related to a limited responsiveness to influenza vaccination and a higher inflammatory status in a cohort of elderly people.

[Increased infections by herpes simplex virus type 1 and polymicrobials of the genital tract, in the general population of a Spanish middle city]. / Aumento de las infecciones por virus del herpes simple tipo 1 y polimicrobianas del aparato genital, en la población general de una ciudad media española.

OBJECTIVE: Infections by genitopathogens are a frequent reason for consultation in Primary Health Care and in the specialties of Infectious Diseases, Urology, Gynecology, and Dermatology. The most common causes are opportunistic microorganisms and responsible for sexually transmitted infections associated with unprotected sex. The objective is to determine the microorganisms that cause these infections in patients treated at the Hospital Universitario Virgen de las Nieves in Granada and Neisseria gonorrhoeae susceptibility to antibiotics. METHODS: A transversal-descriptive and retrospective study was carried out, which included the results issued, between January 2018 and December 2019, in the Microbiology Laboratory from all the episodes studied using standardized working procedures. RESULTS: The most frequently detected microorganisms were Gardnerella vaginalis (23.81%) followed by Candida spp. (20.9%), especially in females, and N. gonorrhoeae (11.36%) and Ureaplasma urealyticum (11.99%), in males. Many times, they were presented in combination. Regarding herpes simplex viruses, infection by both species had a similar prevalence (50%) in males, while type 1 was more prevalent (76.52%) in females. The most active antibiotics against N. gonorrhoeae were cefotaxime (98%) and cefixime (100%). Tetracycline (39.02%) a poorly active antibiotic. CONCLUSIONS: The most frequent pathogens corresponded to those that usually caused infections in females, although N. gonorrhoeae was the most frequent in males and mixed infections are not an accidental finding. HSV-1 infections were more frequent than HSV-2, confirming the trend of a change in the epidemiology of genital herpes.

[Dandy-Walker malformation. The CRIT protocol. A propos of a case]. / Malformación de Dandy-Walker. Protocolo de intervención en CRIT. A propósito de un caso.

Dandy-Walker malformation is characterised by hypoplasia of the cerebellar vermis and cystic dilation of the fourth ventricle. Some of the clinical manifestations of this malformation are hydrocephalus, delayed motor development, hypotonia, and ataxia. Treatment aims to improve the individual's overall functioning and enhance quality of life through a multidisciplinary team. This case report describes the outcome of a patient diagnosed with Dandy-Walker malformation, after the intervention of the medical model at Centro de Rehabilitación e Inclusión Infantil Teletón Guanajuato (CRIT), which focuses on rehabilitating the patient and the family, covering each of the areas encompassing the patient.

Limitation of therapeutic effort in patients hospitalised in departments of internal medicine. / Limitación del esfuerzo terapéutico en pacientes hospitalizados en servicios de medicina interna.

INTRODUCTION: There is little information on the limitation of therapeutic effort (LTE) in patients admitted to hospital internal medicine units. OBJECTIVES: To describe the indicated LTE regimens in the departments of internal medicine and the characteristics of the patients who undergo them. PATIENTS AND METHODS: An observational, descriptive retrospective study was conducted on 4 hospitals of the Community of Madrid. The study collected demographic and comorbidity data and the LTE orders prescribed for all patients who died during a period of 6 months. RESULTS: The study included 382 patients with a mean age of 85±10 years; 204 were women (53.4%) and 222 (58.1%) came from their homes. Some 51.1% of the patients were terminal, 43.2% had moderate to severe dementia, and 95.5% presented at least moderate comorbidity. Some type of LTE was performed in 318 patients (83.7%); the most common orders were "No cardiopulmonary resuscitation" (292 patients, 76.4%; 95% CI 72.1-80.8), "Do not use aggressive measures" (113 patients, 16.4%; 95% CI 13.7-19.4) and "Do not transfer to an intensive care unit" (102 cases, 14.8%, 95% CI 12.3-17.7). Some type of LTE was performed in 318 patients (83.7%); the most common orders were "No cardiopulmonary resuscitation" (292 patients, 76.4%; 95% CI 72.1-80.8), "Do not use aggressive measures" (113 patients, 16.4%; 95% CI 13.7-19.4) and "Do not transfer to an intensive care unit" (102 cases, 14.8%, 95% CI 12.3-17.7). CONCLUSIONS: LTE is common among patients who die in Internal Medicine. The most widely used regimens were "No CPR" and the unspecific statement "Do not use aggressive measures". The patients were elderly and had significant comorbidity, terminal illness and advanced dementia.

Oxidative addition of allylic carbonates to palladium(0) complexes: reversibility and isomerization

The oxidative addition of a cyclic allylic carbonate to the palladium(0) complex generated from a [Pd(dba)2]+2 PPh3 mixture affords a cationic pi-allylpalladium(II) complex with the alkyl carbonate as the counter-anion. This reaction is reversible and proceeds with isomerization of the allylic carbonate at the allylic position. The equilibrium constant has been determined in DMF. The influence of the precursor of the palladium(0) is discussed.

[Severe caustic injuries of the esophagus: when to replace the esophagus]. / Causticaciones esofágicas graves: cuándo decidir prescindir del esófago.

From June 1985 to May 1998, 20 patients have been treated in our hospital by esophageal dilatations due to serious esophageal caustic stenosis. A retrospective analysis of these 20 patients was performed, evaluating age, sex, causative agent, number and time of dilatations, iatrogenic esophageal perforations, gastroesophageal reflux (GER) and psychological and social consequences. Follow-up has ranged from 1.5 to 14.5 years (mean = 8.07 years). The mean age at the time of accidental swallow was 42.2 months. The causative agent was dishwashing detergent in 11 patients and caustic soda in 9 patients. The mean of dilatations needed was 19.1 and the average duration of dilatations was 24.7 months. Seven patients had esophageal perforation during dilatation and none of them required surgical treatment. All patients had improvement of swallowing and an easier esophageal dilatation afterwards. Oral contrast studies demonstrated distortion of the esophago-gastric junction in the majority of patients. Ten patients were screened for GER with 24-hour esophageal pH monitoring in the first year postburn and it was pathological in 5. Antireflux surgery was carried out in 4 patients with a conspicuous improvement following surgery. Dilatations proved successful in 16 patients in less than two years, and in more than two years in 3 patients. The authors conclude that the majority of children with esophageal caustic stenosis can be managed successfully by esophageal dilatations, even with dilatation periods longer than two years or when an esophageal perforation occur during the procedure. The paramount importance of early diagnosis and treatment of GER is stressed.

[Percutaneous tenotomy and aponeurotomy (PTA) for knee flexor contracture in children with spastic cerebral palsy]. / Tenotomía y aponeurotomía percutánea (TAP) para la contractura de los flexores de rodilla en niños con parálisis cerebral espástica.

UNLABELLED: Knee flexor muscle contracture is frequent in patients with spastic cerebral palsy. The purpose of the study was to determine whether percutaneous tenotomy and aponeurotomy may decrease knee flexor contracture in children with spastic cerebral palsy. MATERIAL AND METHODS: A prospective study of consecutive cases was conducted from January to December 2009 in 24 children with a diagnosis of moderate to severe spastic cerebral palsy who had knee flexor contracture with a popliteal angle > or = 45 degrees and a gross motor function classification scale of 4 or 5; they underwent percutaneous tenotomy and aponeurotomy surgery and were followed-up for 24 months. Variance analysis with a factorial design was used for data analysis. RESULTS: The mean popliteal angle was 83.48 degrees preoperatively and 27.30 degrees by the end of the follow-up, with an improvement of 56.18 degrees (p < 0.01). Statistically significant differences were found in all measurements comparing them with the baseline values. DISCUSSION: Percutaneous aponeurotomy of knee flexor muscles is described. Compared to other procedures it provides the benefits of minimally invasive surgery, mild postoperative pain, short hospital stay -without using immobilization during the entire process- and children returned to their therapy program within five days. CONCLUSION: Percutaneous tenotomy and aponeurotomy of knee flexors was shown to be a good alternative for the treatment of knee flexor contracture in patients with spastic cerebral palsy.

Rate and mechanism of the reversible formation of cationic (eta3-allyl)-palladium complexes in the oxidative addition of allylic acetate to palladium(0) complexes ligated by diphosphanes.

The oxidative addition of the allylic acetate, CH2=CH-CH2-OAc, to the palladium(o) complex [Pd0(P,P)], generated from the reaction of [Pd(dba)2, with one equivalent of P,P (P,P = dppb = 1,4-bis(diphenylphosphanyl)butane, and P,P = dppf = 1,1'-bis(diphenylphosphanyl)ferrocene), gives a cationic (eta3-allyl)palladium(II) complex, [(eta3-C3H5)Pd(P,P)+]. with AcO as the counter anion. This reaction is reversible and proceeds through two successive equilibria. The overall equilibrium constants have been determined in DMF. Compared with PPh3, the overall equilibrium lies more in favor of the cationic (eta3-allyl)palladium(II) complex when bidentate P,P ligands are considered in the order: dppb > dppf > PPh3. The reaction proceeds via a neutral intermediate complex [(eta2-CH=CH-CHCH2-OAc)Pd0(P,P)], which has been kinetically detected. The rate constants of the successive steps have been determined in DMF by UV spectroscopy and conductivity measurements. The overall complexation step of the Pd0 by the allylic acetate C=C bond is faster than the oxidative addition/ionization step which gives the cationic (eta3-allyl)palladium(II) complex.

Rapid detection of respiratory viruses by shell vial assay using simultaneous culture of HEp-2, LLC-MK2, and MDCK cells in a single vial.

A shell vial assay with simultaneous culture of HEp-2, LLC-MK2, and MDCK cell lines in a single tube (CoHLM SV assay) was compared with traditional tube culture (TC) for the detection of the main respiratory viruses in 358 nasal wash specimens. A total of 170 strains were isolated from 168 virus-positive samples. A total of 94. 1% of the strains (160 strains; 128 respiratory syncytial viruses and 32 other viruses) were detected by the CoHLM SV assay in 48 h, whereas 98.2% of the strains (167 strains; 132 respiratory syncytial viruses and 35 other viruses) were detected by TC in a mean time of 6 days. The CoHLM SV assay may be useful for the rapid detection of respiratory viruses.