Use of a recombinant positive control in the diagnostic of canine Ehrlichiosis from 16sRNA gen of Ehrlichia canis in Mexico City.

Ehrlichia canis has gained importance over the years as a zoonotic bacterium, nevertheless in Mexico is unknown the extent of the problem in animals and public health. The country had a few studies carried out locally using serology and molecular tests as diagnostic methods. Ehrlichiosis is not considered endemic in the central valley of Mexico, because the climatic conditions in the region have not allowed the vector (Rhipicephalus sanguineus) to establish itself adequately, therefore, diagnosis is not used in clinical practice in this area. A nested PCR (nPCR) offers rapid results with high sensitivity and specificity regardless of cost. The use of a recombinant positive control provides the advantage of timely diagnosis, follow-up treatment and allows the clinician to decide. In this work, the nPCR reported by Wen et al. (J Clin Microbiol 35(7):1852-2185, 1997) was used for the diagnosis of E. canis by modifying the reaction conditions to improve the detection of the test. We constructed a recombinant positive control to nPCR as diagnostic technique for E. canis, also we modified the reaction conditions to improve detection of the test which allowed the diagnosis of E. canis in dogs in the Mexican Republic using 53 samples from dogs with positive serological diagnosis of Ehrlichiosis, some of them from the valley of Mexico. Currently, this nPCR is offered to public at the Faculty of Veterinary Medicine and Zootechnics of the National Autonomous University of Mexico at an accessible cost and allows to begin to generate epidemiological information to know distribution of the bacterium.

Effect of size disparity on the structure and dynamics of the small component in concentrated binary colloidal mixtures.

We determined, using confocal microscopy, the structure and dynamics of the small component in concentrated binary colloidal mixtures with moderate and large size ratios and different compositions of Polymethyl methacrylate particles. We show that when increasing the content of small spheres at fixed total volume fraction, a transition in the local environment of the small particles is observed, from a mixed environment of other small and large particles to a local environment of only small particles. The transition is rather abrupt for moderate size ratios, while it becomes particularly broad for large size ratios. This can be associated with the improved ability of the small particles to pack in between the large particles for larger size ratios. The dynamics reflect the transition with an increase of the mobility observed at intermediate mixing. This increase becomes particularly pronounced for large size ratios, leading to diffusive dynamics of the small particles, in agreement with predictions of theories of the glass transition in binary hard-sphere mixtures. The composition at which the fastest dynamics are observed is apparently independent of the size ratio.

Microliter viscometry using a bright-field microscope: η-DDM.

The rheological properties of a medium can be inferred from the Brownian motion of colloidal tracer particles using the microrheology procedure. The tracer motion can be characterized by the mean-squared displacement (MSD). It can be calculated from the intermediate scattering function determined by Differential Dynamic Microscopy (DDM). Here we show that DDM together with the empirical Cox-Merz rule is particularly suited to measure the steady-shear viscosity, i.e. the viscosity towards zero frequency, due to its ability to provide reliable information on long time and length scales and hence small frequencies. This method, η-DDM, is tested and illustrated using three different systems: Newtonian fluids (glycerol-water mixtures), colloidal suspensions (protein samples) and a viscoelastic polymer solution (aqueous poly(ethylene oxide) solution). These tests show that common lab equipment, namely a bright-field optical microscope, can be used as a convenient and reliable microliter viscometer. Because η-DDM requires much smaller sample volumes than classical rheometry, only a few microliters, it is particularly useful for biological and soft matter systems.

Investigation of moderately turbid suspensions by heterodyne near field scattering.

Light scattering has proven to be a very powerful technique to characterize soft matter systems. However, many samples are turbid and hence suffer from multiple scattering which can affect the signal considerably. Multiple scattering can be reduced by diluting the sample or changing the solvent, but often this alters the sample and hence is precluded. Here we study the dynamics of a model system. In particular, we investigate the effects of moderate multiple scattering on small-angle heterodyne near field scattering (HNFS). Varying the particle concentration and size we change the degree of multiple scattering, which is quantified by the transmission of light. In dependence of the degree of multiple scattering, we analyze the statistical properties of the HNFS signal, which is the difference between two intensity patterns separated by a delay time. The distribution of intensity differences follows a Gaussian distribution if single scattering dominates and a Laplace distribution in the presence of extreme multiple scattering. We also investigate the effects of multiple scattering on the measured intermediate scattering function and the hydrodynamic radius of the particles. Reliable data are obtained for sample transmissions down to about 0.7. This is confirmed by a comparison with results from a far field cross-correlation instrument that suppresses multiple scattering contributions. Therefore, HNFS represents a technically simple but powerful method to investigate samples that are moderately multiple scattering.

Analysis of PTPN22, ZFAT and MYO9B polymorphisms in Turner Syndrome and risk of autoimmune disease.

Turner syndrome (TS) is one of the most common sexual chromosome abnormalities and is clearly associated with an increased risk of autoimmune diseases, particularly thyroid disease and coeliac disease (CD). Single-nucleotide polymorphism analyses have been shown to provide correlative evidence that specific genes are associated with autoimmune disease. Our aim was to study the functional polymorphic variants of PTPN22 and ZFAT in relation to thyroid disease and those of MYO9B in relation to CD. A cross-sectional comparative analysis was performed on Mexican mestizo patients with TS and age-matched healthy females. Our data showed that PTPN22 C1858T (considered a risk variant) is not associated with TS (X2  = 3.50, p = .61, and OR = 0.33 [95% CI = 0.10-1.10]). Also, ZFAT was not associated with TS (X2  = 1.2, p = .28, and OR = 1.22 [95% CI = 0.84-1.79]). However, for the first time, rs2305767 MYO9B was revealed to have a strong association with TS (X2  = 58.6, p = .0001, and OR = 10.44 [95% C = 5.51-19.80]), supporting a high level of predisposition to CD among TS patients. This report addresses additional data regarding the polymorphic variants associated with autoimmune disease, one of the most common complications in TS.

Brainstem and Autonomic Nervous System Dysfunction: A Neurosurgical Point of View.

Central autonomic control nuclei and pathways are mainly integrated within the brainstem, especially in the medulla oblongata. Lesions within these structures can lead to central dysautonomia.Central autonomic control structures can be damaged by tumors, during surgery, or by other neurosurgical pathologies. These may elicit clinical or subclinical autonomic complications that can constitute a serious clinical problem.The authors present a broad review of the central autonomic nervous system, its possible dysfunctions, and the relation between neurosurgery and this "not-well-known system". Preliminary results of an autonomic study of brainstem lesions that is currently being carried out by the authors are also shown.

Opinion survey on proposals for improving code stroke in Murcia Health District V, 2014. / Encuesta de opinión sobre propuestas de mejora del código ictus en el Área V de la Región de Murcia, 2014.

INTRODUCTION: Stroke is a time-dependent neurological disease. Health District V in the Murcia Health System has certain demographic and geographical characteristics that make it necessary to create specific improvement strategies to ensure proper functioning of code stroke (CS). The study objectives were to assess local professionals' opinions about code stroke activation and procedure, and to share these suggestions with the regional multidisciplinary group for code stroke. SUBJECTS AND METHOD: This cross-sectional and descriptive study used the Delphi technique to develop a questionnaire for doctors and nurses working at all care levels in Area V. An anonymous electronic survey was sent to 154 professionals. The analysis was performed using the SWOT method (Strengths, Weaknesses, Opportunities, and Threats). RESULTS: Researchers collected 51 questionnaires. The main proposals were providing training, promoting communication with the neurologist, overcoming physical distances, using diagnostic imaging tests, motivating professionals, and raising awareness in the general population. CONCLUSIONS: Most of the interventions proposed by the participants have been listed in published literature. These improvement proposals were forwarded to the Regional Code Stroke Improvement Group.

Urotensinergic system genes in experimental subarachnoid hemorrhage.

OBJECTIVE: Cerebral vasospasm, one of the main complications of subarachnoid hemorrhage (SAH), is characterized by arterial constriction and mainly occurs from day 4 until the second week after the event. Urotensin-II (U-II) has been described as the most potent vasoconstrictor peptide in mammals. An analysis is made of the serum U-II concentrations and mRNA expression levels of U-II, urotensin related peptide (URP) and urotensin receptor (UT) genes in an experimental murine model of SAH. DESIGN: An experimental study was carried out. SETTING: Experimental operating room of the Biomedicine Institute of Seville (IBiS), Virgen del Rocío University Hospital (Seville, Spain). PARTICIPANTS: 96 Wistar rats: 74 SAH and 22 sham intervention animals. INTERVENTIONS: Day 1: blood sampling, followed by the percutaneous injection of 100µl saline (sham) or blood (SAH) into the subarachnoid space. Day 5: blood sampling, followed by sacrifice of the animals. MAIN VARIABLES OF INTEREST: Weight, early mortality, serum U-II levels, mRNA values for U-II, URP and UT. RESULTS: Serum U-II levels increased in the SAH group from day 1 (0.62pg/mL [IQR 0.36-1.08]) to day 5 (0.74pg/mL [IQR 0.39-1.43]) (p<0.05), though not in the sham group (0.56pg/mL [IQR 0.06-0.83] day 1; 0.37pg/mL [IQR 0.23-0.62] day 5; p=0.959). Between-group differences were found on day 5 (p<0.05). The ROC analysis showed that the day 5 serum U-II levels (AUC=0.691), URP mRNA (AUC=0.706) and UT mRNA (AUC=0.713) could discriminate between sham and SAH rats. The normal serum U-II concentration range in rats was 0.56pg/mL (IQR 0.06-0.83). CONCLUSION: The urotensinergic system is upregulated on day 5 in an experimental model of SAH.

Impact of Vitiligo on Quality of Life. / Impacto del vitíligo en la calidad de vida.

INTRODUCTION AND OBJECTIVE: Vitiligo is a chronic autoimmune skin disease caused by the destruction of melanocytes. Although quality of life (QOL) in vitiligo has been studied in different countries, it has not yet been investigated in Mexico. The aim of this study was to assess the QOL of Mexican patients with vitiligo. MATERIAL AND METHOD: We conducted a cross-sectional study at the research unit of Centro Dermatológico Dr. Ladislao de la Pascua in Mexico City. We included adults with vitiligo and excluded those with other pigmentation disorders or a neurological or psychiatric disorder. Patients on psychoactive medications were also excluded. All the patients were administered the Dermatology Life Quality Index (DLQI), a vitiligo-specific quality of life instrument (the VitiQoL), and the Beck Depression and Anxiety Inventories. RESULTS: We studied 150 patients with vitiligo (103 women [68.7%] and 47 men [31.3%]). The median (interquartile range) age was 38 (20) years. The mean (SD) scores on the DLQI and VitiQoL were 5.2 (5.4) and 32.1 (22.7) out of total possible scores of 30 and 90, respectively. The correlation between questionnaire scores was 0.675 (P<.001). Patients with genital involvement scored significantly worse on the VitiQoL than those without lesions in this area (43.95 [28.4]) vs. 28.98 [20.08], P<.001). The prevalence of depression and anxiety was 34% and 60%, respectively. CONCLUSION: Vitiligo has a minimal impact on the QOL of our patients. QOL was worse in patients with genital lesions.

Fluorescence and Textural Characterization of Ortho-Amine Tetraphenylporphyrin Covalently Bonded to Organo-Modified Silica Xerogels.

Most of the studies performed with porphyrins involve these species functionalized with peripheral substituents lying on the same macrocyclic molecular plane. The main objective of this work deals with the successful preservation and optimization of the fluorescence of a uncommonly used porphyrin species, i.e. tetrakis-(ortho-amino-phenyl)-porphyrin; a molecule with substituents localized not only at one but at both sides of its molecular plane. In cases like this, it must be stressed that fluorescence can only be partially preserved; nevertheless, intense fluorescence can still be reached by following a twofold functionalization strategy involving: (i) the bonding of substituted macrocycles to the pore walls of (ii) organo-modified silica monoliths synthesized by the sol-gel method. The analysis of both absorption and emission UV spectra evidenced a radiation energy transfer taking place between the porphyrin and the host silica matrix. Our results showed that the adequate displaying of the optical properties of macrocyclic species trapped in SiO2 xerogels depend on the polarity existing inside the pores, a property which can be tuned up through the adequate selection of organic groups used to modify the surface of the pore cavities. Additionally, the pore widths attained in the final xerogels can vary depending on the identity of the organic groups attached to the network. All these facts finally demonstrated that, even if using inefficient surface functionalization species, such as ortho-substituted tetraphenylporphyrins, it is still possible to modulate the pore shape, pore size, and physicochemical environment created around the trapped macrocycles. The most important aspect related to this research deals with the fact that the developed methodology offers a real possibility of controlling both the textural and morphological characteristics of a new kind of hybrid porous materials and to optimize the physicochemical properties of diverse active molecules trapped inside the pores of these materials.

[Social representations of public health among healthcare workers within the French Armed Forces]. / Les représentations sociales de la santé publique chez les professionnels de santé dans les armées françaises.

BACKGROUND: Public health is a multidisciplinary activity whose fields of action are acquiring an increasingly broad. The Service de santé des armées (SSA) has always had a culture of public health problems thanks to doctors specialized in the treatment of major diseases. Often involved in public health activities, health professionals nevertheless have a fragmented vision. The objective was to describe the social representations of public health of military healthcare workers. METHOD: The responders were doctors, nurses, veterinarians and pharmacists practicing in different areas of SSA (caregivers, administrators, policy makers) and were interviewed by telephone. A question of spontaneous evocation on the representations of public health in the army was asked. The overall lexical analysis was performed according to the method of rank-frequency. Categorical analysis was conducted to better understand the whole lexical field use. RESULTS: There were 90 responders. The most salient terms were "prevention, epidemiological surveillance and vaccination". The categorical analysis showed that doctors used a lexical field primarily focused on diseases and risk behaviors, nurses on the specifics of military surveillance and policy makers about the cross-discipline. CONCLUSION: Public health in the army is mainly represented by epidemiological surveillance and prevention. Given the non-mentioned fields, a strengthening of communication on the current challenges of public health would probably improve adherence of healthcare professionals since public health takes on more and more importance in the development of the national health policy and management of health crises.

Chlorophyll a Covalently Bonded to Organo-Modified Translucent Silica Xerogels: Optimizing Fluorescence and Maximum Loading.

Chlorophyll is a pyrrolic pigment with important optical properties, which is the reason it has been studied for many years. Recently, interest has been rising with respect to this molecule because of its outstanding physicochemical properties, particularly applicable to the design and development of luminescent materials, hybrid sensor systems, and photodynamic therapy devices for the treatment of cancer cells and bacteria. More recently, our research group has been finding evidence for the possibility of preserving these important properties of substrates containing chlorophyll covalently incorporated within solid pore matrices, such as SiO2, TiO2 or ZrO2 synthesized through the sol-gel process. In this work, we study the optical properties of silica xerogels organo-modified on their surface with allyl and phenyl groups and containing different concentrations of chlorophyll bonded to the pore walls, in order to optimize the fluorescence that these macrocyclic species displays in solution. The intention of this investigation was to determine the maximum chlorophyll a concentration at which this molecule can be trapped inside the pores of a given xerogel and to ascertain if this pigment remains trapped as a monomer, a dimer, or aggregate. Allyl and phenyl groups were deposited on the surface of xerogels in view of their important effects on the stability of the molecule, as well as over the fluorescence emission of chlorophyll; however, these organic groups allow the trapping of either chlorophyll a monomers or dimers. The determination of the above parameters allows finding the most adequate systems for subsequent in vitro or in vivo studies. The characterization of the obtained xerogels was performed through spectroscopic absorption, emission and excitation spectra. These hybrid systems can be employed as mimics of natural systems; the entrapment of chlorophyll inside pore matrices indicates that it is possible to exploit some of the most physicochemical properties of trapped chlorophyll for diverse technological applications. The data herein collected suggest the possibility of applying the developed methodology to other active, captive molecules in order to synthesize new hybrid materials with optimized properties, suitable to be applied in diverse technological fields.

Transoral robotic surgery of the central skull base: preclinical investigations.

In this study we explored possible applications of the da Vinci system in approaching the skull base at optic chiasm level on two cryopreserved cadavers, using an entirely transoral robotic technique (TORS). We used a standard 12 mm endoscopy and 8 mm terminals. Bone drilling was performed manually. The da Vinci system is equipped with very good illumination and 3D viewing, thus providing excellent vision and great maneuverability even in the less accessible areas of the skull. Our experience demonstrates that an entirely transoral skull base robotic approach to this complex anatomical region has many advantages as compared to traditional techniques.

[Laparoscopic cholecystectomy: what we can learn about]. / Colecistectomía laparoscópica en niños y adultos: lo que podemos aprender.

BACKGROUND: Laparoscopic cholecystectomy (LC) is a very usual procedure within adult population, but not as frequent in childhood. The aim of this study was to assess the outcome of LC in children compared with those performed in adulthood. MATERIALS AND METHODS: We reviewed 39 consecutive patients who underwent LC between 2003 and 2013 at our Department and a similar sample of patients from 18 to 40 years of age from the General Surgery Department. RESULTS: 39 children and 40 adults fulfilled criteria to be included in the study. The most frequent indication was cholelithiasis in both groups. The mean operating time was significantly higher among children (127 min, adults 71 min, p < 0.01) but we didn't find differences neither in conversion nor in complication rates (children 5% and 7.7%, adults 2.5% and 15% respectively). In regard to preoperative factors, only male gender was correlated to a higher complication rate (p 0.037). On the other hand we found out that, in absence of complications, both the average length of stay (children 2.1 days, adults 0.5 days) and mean time to first feeding (children 21 hours, adults 8 hours) were significantly higher among children (p < 0.01). CONCLUSIONS: 1) LC in childhood is a safe procedure that does not imply more morbidity than the same intervention in adults, even though a more prolonged operating time. 2) We believe that our longer hospital stay is due to certain lack of confidence with the technique and, in the future, the trend should be bent on encouraging a shorter time to first feeding and an earlier discharge.

[An experimental model of mass-type brain damage in the rat: expression of brain damage based on neurospecific enolase and protein S100B]. / Modelo experimental de lesión cerebral tipo masa en rata: expresión del daño cerebral mediante enolasa neuroespecífica y proteína S100B.

OBJECTIVE: To determine whether a model of transient mass-type brain damage (MTBD) in the rat produces early release of neurospecific enolase (NSE) and protein S100B in peripheral blood, as an expression of the induced brain injury. DESIGN: An experimental study with a control group. SETTING: Experimental operating room of the Institute of Biomedicine (IBiS) of Virgen del Rocío University Hospital (Seville, Spain). PARTICIPANTS: Fourteen adult Wistar rats. INTERVENTIONS: Blood was sampled at baseline, followed by: MTBD group, a trephine perforation was used to insert and inflate the balloon of a catheter at a rate of 500 µl/20 sec, followed by 4 blood extractions every 20 min. Control group, the same procedure as before was carried out, though without trephine perforation. PRIMARY STUDY VARIABLES: Weight, early mortality, serum NSE and S100B concentration. RESULTS: Differences in NSE and S100B concentration were observed over time within the MTBD group (P<.001), though not so in the control group. With the exception of the baseline determination, differences were observed between the two groups in terms of the mean NSE and S100B values. Following MTBD, NSE and S100B progressively increased at all measurement timepoints, with r=0.765; P=.001 and r=0.628; P=.001, respectively. In contrast, the control group showed no such correlation for either biomarker. CONCLUSIONS: Serum NSE and S100B concentrations offer an early indication of brain injury affecting the gray and white matter in an experimental model of mass-type MTBD in the rat.

Design and validation of a questionnaire for measuring perceived risk of skin cancer.

INTRODUCTION: A perceived risk of cancer encourages preventive behavior while the lack of such a perception is a barrier to risk reduction. There are no instruments in Spanish to measure this perceived risk and thus quantify response to interventions for preventing this disease at a population level. The aim of this study was to design and validate a self-administered questionnaire for measuring the perceived risk of skin cancer. MATERIAL AND METHODS: A self-administered questionnaire with a visual Likert-type scale was designed based on the results of the analysis of the content of a survey performed in 100 patients in the Dr. Ladislao de la Pascua Skin Clinic, Distrito Federal México, Mexico. Subsequently, the questionnaire was administered to a sample of 359 adult patients who attended the clinic for the first time. As no gold standard exists for measuring the perceived risk of skin cancer, the construct was validated through factor analysis. RESULTS: The final questionnaire had 18 items. The internal consistency measured with Cronbach α was 0.824 overall. In the factor analysis, 4 factors (denoted as affective, behavioral, severity, and susceptibility) and an indicator of risk accounted for 65.133% of the variance. CONCLUSIONS: The psychometric properties of the scale were appropriate for measuring the perception of risk in adult patients (aged 18 years or more) who attended the dermatology clinic.

[Translated article] Mechanical resistance of polylactic acid bone matrices developed by 3D printing for the reconstruction of bone defects.

INTRODUCTION: Bone defects are one of the main limitations in orthopaedic surgery and traumatology. For this reason, multiple bone replacement systems have been developed, either by prosthetic implant or by substitution with osteoforming substances, whose limitations are their survival and lack of structurality, respectively. The objective of this work is the generation of a new material for the creation of biologically active structures that have sufficient tensile strength to maintain the structure during remodelling. MATERIAL AND METHODS: A new filament based on the fusion of natural polylactide acid (PLA) powder was designed for the generation of pieces by means of fused deposition modelling (FDM) on which to carry out tensile mechanical tests of osteosynthesis material. A total of 13 groups with different cortical thickness, filling and layer height were carried out, with 10 tensile tests in each group, defining the tensile breaking limit for each group. The regression lines for each group and their mechanical resistance to traction on the filament used were determined. RESULTS: The filament ratio per contact surface unit with the osteosynthesis used was the main determinant of the mechanical resistance to traction, either at the expense of the increase in cortical thickness or by the increase in the percentage of cancellous bone filling. Layer height had a minor effect on tensile strength. The regression value was high for cortical thickness and cancellous filling, being elements with a predictable biomechanical behaviour. CONCLUSIONS: The new methodology allows the creation of personalised neutral and implantable PLA bone matrices for the reconstruction of large bone defects by means of 3D printing by FDM with a mechanical resistance to traction greater than that of current biological support structures.

Antimicrobial evaluation of silver nanoparticles using extracts of Crescentia cujete L.

New natural reducing agents with a lower negative impact on the environment and with a high antimicrobial potential are required for the process of obtaining silver nanoparticles through the chemical reduction method. The use of plant extracts can be a fast track in the formation of nanoparticles. In this case, organic compounds such as terpenes, flavonoids, enzymes, proteins, and cofactors present in plants act as reducing agents for nanomaterials. This research evaluated the antimicrobial property of silver nanoparticles from extracts of Crescentia cujete L. The presence of quercetin (flavonoid) was determined by high-performance liquid chromatography (HPLC); the production of silver nanoparticles (AgNPs) was established by green synthesis; the size and morphology of the nanomaterials were evaluated by scanning electron microscope (SEM). The antimicrobial capacity was studied by two analysis methods: modified culture medium and surface seeding. The presence of quercetin (26.55 mg L-1) in the crude extract of Crescentia cujete L., identified by HPLC, was evidenced. Nanoparticle formation was spherical, with an average size of 250 ± 3 and 460 ± 6 nm. Microbiological cultures with treatment showed 94% microbial inhibition. It was concluded that the Crescentia cujete L., leaves shoed an acceptable concentration of quercetin to be used as a useful adjuvant to enhance the reduction of NPs synthesis. The nanoparticles produced by green synthesis proved to have a positive effect to combat pathogenic microorganisms.