Co-occurrence and seasonal and environmental distributions of the sandflies Lutzomyia longipalpis and Nyssomyia whitmani in the city of Puerto Iguazú, northeastern Argentina.

The aim of this work was to study the distribution of Phlebotominae (Diptera: Psycodidade) abundance in time and space in an area in northeastern Argentina with vector transmission of visceral and tegumentary leishmaniasis. For this, 51 households were selected using a 'worst scenario' criterion where one light trap was set during two consecutive nights in peridomiciles in the transitions between the four seasons, and the environment was surveyed simultaneously. The relationships of phlebotomine assemblage structure and the most abundant species with seasonality and environmental variables were evaluated using a canonical correspondence analysis and generalized linear mixed models, respectively. A total of 5110 individuals were captured. Lutzomyia longipalpis (Lutz & Neiva, 1912) and Nyssomyia whitmani (Antunes & Coutinho, 1939) were the most abundant species captured in all samplings (98.3% of the total capture). The period of highest abundance of Lu. longipalpis was early autumn, and it was distributed in the most urbanized areas. Nyssomyia whitmani occupied mainly the less urbanized areas, showing peaks of abundance in early spring and summer. Other species were captured in low numbers and showed seasonal-spatial variations similar to those of Ny. whitmani. We confirmed Leishmania spp. vector persistence throughout the year in spatial patches of high abundance even during the less favorable season.

Is the snail shell repair process really influenced by eggshell membrane as a template of foreign scaffold?

Biominerals are inorganic-organic hybrid composites formed via self-assembled bottom up processes under mild conditions. Biominerals show interesting physical properties, controlled hierarchical structures and robust remodeling or repair mechanisms. Biological processes associated with biominerals remain to be developed into practical engineering processes. Therefore, the formation of biominerals is inspiring for the design of materials, especially those fabricated at ambient temperatures. The study described herein involves the influence of chicken outer eggshell membrane on the type of calcium carbonate (CaCO3) polymorph deposited on the shell of the land snail Helix aspersa during the repair process after an injury. A piece of snail shell was removed by perforating a hole from the largest body whorl. The operated area was left either uncovered or covered with either a thermoplastic flexible polyolefin-based film Parafilm® or a piece of chicken eggshell membrane. The repaired shells of control and experimental animals were analyzed using SEM, EDS, Raman and FTIR spectroscopies. We found that in the presence of eggshell membrane, the polymorph deposited on the substratum during the first hours resembles calcite, the polymorph present in eggshell normal formation, but at 24 and 48h, when snail mantle cells produced their normal organic matrix (mainly ß-chitin plus proteins and proteoglycans), the polymorph deposited is aragonite, the characteristic polymorph of Helix shell. Therefore, the eggshell membrane influences the type of polymorph, but only in the initial stages of biomineral deposition, before an organic matrix layer is deposited by the snail.

Austromegabalanus psittacus barnacle shell structure and proteoglycan localization and functionality.

Comparative analyzes of biomineralization models have being crucial for the understanding of the functional properties of biominerals and the elucidation of the processes through which biomacromolecules control the synthesis and structural organization of inorganic mineral-based biomaterials. Among calcium carbonate-containing bioceramics, egg, mollusk and echinoderm shells, and crustacean carapaces, have being fairly well characterized. However, Thoraceca barnacles, although being crustacea, showing molting cycle, build a quite stable and heavily mineralized shell that completely surround the animal, which is for life firmly cemented to the substratum. This makes barnacles an interesting model for studying processes of biomineralization. Here we studied the main microstructural and ultrastructural features of Austromegabalanus psittacus barnacle shell, characterize the occurrence of specific proteoglycans (keratan-, dermatan- and chondroitin-6-sulfate proteoglycans) in different soluble and insoluble organic fractions extracted from the shell, and tested them for their ability to crystallize calcium carbonate in vitro. Our results indicate that, in the barnacle model, proteoglycans are good candidates for the modification of the calcite crystal morphology, although the cooperative effect of some additional proteins in the shell could not be excluded.

Self-contamination following removal of two personal protective equipment suits: a randomized, controlled, crossover simulation trial.

BACKGROUND: Healthcare personnel are often at high risk of contamination when participating in airway management and other aerosol-generating procedures. AIM: To explore the differences in self-contamination after removal of gown and coverall personal protective equipment (PPE) using an ultraviolet-fluorescent solution. METHODS: This prospective, randomized, controlled crossover trial was set in a third-level university health centre in Buenos Aires, Argentina between August and October 2020. The study included 60 anaesthesia personnel volunteers, and no participants were excluded from the study. A two-period/two-intervention design was chosen; each intervention comprised audio-guided placement of PPE, full-body spraying of fluorescent solution, audio-guided removal of PPE, and self-contamination assessment through ultraviolet light scanning. The primary outcome was the mean within-participant difference (any traces) between PPE suits. Statistical significance was tested using t-tests for paired data. The allocation ratio was 25/35 (gown followed by coverall/coverall followed by gown). FINDINGS: Self-contamination after removal of coveralls was greater than that after removal of gowns, with a mean within-participant difference of 11.45 traces (95% confidence interval 8.26-14.635; P<0.001). Significant differences were found for the number of self-contaminated body zones, small fluorescent traces and large fluorescent traces. Removal of a gown was associated with a markedly lower risk of self-contamination. CONCLUSIONS: Quick one-step removal of a gown and gloves may reduce self-contamination in the arm/hand area. Fluorescent solutions can help to identify self-contamination and compare outcomes between available PPE suits. Repeated training sessions and enhanced knowledge on self-contamination following removal of PPE are paramount. CLINICAL TRIAL REGISTRATION NUMBER: NCT04763304 (on ClinicalTrials.gov).

[Malignant arterial hypertension (HTAM) with severe systolic dysfunction of the reversible left ventricle]. / Hipertensión arterial maligna (HTAM) con disfunción sistólica grave del ventrículo izquierdo reversible.

Malignant arterial hypertension is still present in current clinical care despite the fact that for more than three decades we have had a wide range of antihypertensive drugs to control high blood pressure. It is essential to know how to detect it in time due to its high risk to life, with poor short-term prognosis if not treated properly. It may present with nonspecific, but potentially serious, clinical symptoms or manifest clinically as a hypertensive emergency accompanied by hypertensive encephalopathy and multi-organ failure. We present a case of a 49-year-old woman, attended in our hospital who had an initial hypertension of 223/170mmHg accompanied by multi-organ failure, who progressed satisfactorily with antihypertensive treatment.

Stress-induced flexibility and individuality in female and male zebra finch distance calls.

Vocal recognition is central to the coordination and organization of behavior in pair-bonding species such as zebra finches. Zebra finches' vocalizations are individualized and support acoustic discrimination processes. Physiological states - such as the ones involved in emotional stress - can modify vocal production and consequently the structure of vocalizations. These modifications might signal the state of the caller but also impair individual recognition processes. This may represent a signaling trade-off, especially in contexts where both pieces of information can be critically important, for example when mates use calls to reunite after social isolation. Here we study the impact of a stress on the individual vocal signature in both female and male zebra finch distance calls. We built a manually curated database of distance calls of several individuals (both females and males) recorded in control and stress conditions. The stress was induced either by social isolation of the bird or using exogenous corticosterone. We developed a machine learning approach to assess the impact of stress on the individual characterization of calls. We show that while calls' spectral structure is significantly modified by stress, it still allows for the correct classification of calls to the caller. Moreover, we also show that the stress-induced modification of calls' structure is not a 'general feature signal' that can be detected as a 'stress' signal regardless of identity. Thus, female and male zebra finch calls' structure show stress-induced flexibility that stays within the range of individual vocal signatures.

[Trapped epidural catheter: reconstruction of computed tomography images]. / Atrapamiento de un catéter epidural. Reconstrucción de imágenes de tomografía computarizada.

A trapped epidural catheter without a knot is a rare complication. During placement of an epidural catheter for analgesia during labor, resistance made it impossible to position the catheter within the epidural space. A second catheter was inserted to provide the required analgesia. When the second catheter was removed, computed tomography (CT) revealed that the tip of the first catheter was close to the the right facet joint space. A second attempt to extract the catheter failed. In light of this situation, the patient was seated with the spine slightly bent to one side, a guidewire was inserted through the catheter lumen, and the catheter and guidewire were gently pulled; the catheter was extracted without causing the tip to break up. Three-dimensional CT reconstruction allowed the catheter tip and characteristics of the joint surfaces to be observed. We discuss protcols and alternative strategies that can be followed when an epidural catheter is difficult to remove, including the most appropriate images to use for guidance.

A prospective international Aspergillus terreus survey: an EFISG, ISHAM and ECMM joint study.

OBJECTIVES: A prospective international multicentre surveillance study was conducted to investigate the prevalence and amphotericin B susceptibility of Aspergillus terreus species complex infections. METHODS: A total of 370 cases from 21 countries were evaluated. RESULTS: The overall prevalence of A. terreus species complex among the investigated patients with mould-positive cultures was 5.2% (370/7116). Amphotericin B MICs ranged from 0.125 to 32 mg/L, (median 8 mg/L). CONCLUSIONS: Aspergillus terreus species complex infections cause a wide spectrum of aspergillosis and the majority of cryptic species display high amphotericin B MICs.

[Hypertensive crisis and sudden change of vision in young patients]. / Crisis hipertensiva y alteración brusca de la visión en el paciente joven.

The sudden change in vision is a medical emergency that must be evaluated immediately to rule out important institutions as systemic vasculitis or ischemic stroke. Its association with hypertensive crisis makes it necessary to rule out accelerated-malignant hypertension, which is accompanied by other retinal disorders (exudates and hemorrhages) and adrenal involvement. Nonarteritic anterior ischemic optic neuropathy (AION) is another entity to consider, as is it not uncommon in the young (12.7% in a series of 848 cases). Its association with hypertension has been described in 32% of cases.

1H-NMR of phosphatidylcholine liposomes at low p2H in the presence of a paramagnetic shift reagent.

1H-NMR spectra of egg phosphatidylcholine liposomes in 2H2O were obtained at several p2H values in the presence of 10 mM PrCl3 added after sonication of the phospholipid. It has been found that as the p2H is lowered below 2, the two distinct signals corresponding to the outer and inner phospholipid trimethylammonium groups which arise by the shifting effect of the paramagnetic cation on the external surface of vesicles, tend to coalesce into a single, high-field peak, at the position corresponding to the internal, non-shifted -N+ (CH3)3 protons. These results can be interpreted to mean that the shifting effect of Pr3+ on phosphatidylcholine NMR spectra, is due to electrostatic interaction between the lanthanide and the ionized group of the lipid. At low p2H, as the phosphodiester becomes protonated, the paramagnetic cation is no longer attracted by the liposome surface and its shifting effect on the phospholipid NMR signals disappears. The plot of the p2H dependence of the chemical shift of the outer trimethylammonium resonance of phosphatidylcholine liposomes with praseodymium ions present only on the outside of vesicles, results in a sigmoidal titration curve with its midpoint at p2H 1.5. In contrast, the inner signal is not affected by p2H. If coalescence of signals is considered as indicative of complete protonation of the phosphate moiety, the value of 1.5 can be taken as the apparent pK for the ionization of that group under the experimental conditions employed, i.e., 10 mM PrCl3 in 2H2O. That the low p2H-induced merging of the signals is reversible, is shown by the reappearance of the two peaks when the p2H of the phospholipid dispersion is raised from 1 to 5.7. Since the recovery of the trimethylammonium signal splitting indicates that Pr3+ has remained excluded from the liposome inner compartment, these experiments also demonstrate that the vesicles have not been disrupted by exposure to such an extreme acidic condition as p2H 1.

Formation of micelles and membrane action of the local anesthetic tetracaine hydrochloride.

The formation of micelles of the local anesthetic tetracaine hydrochloride in aqueous phosphate buffer solution of pH 6.5 and ionic strength (I) 0.10 was examined at 22 degrees C by surface tension and using the fluorescent indicators perylene (peri-dinaphthalene) and 8-anilino-1-naphthalene sulfonic acid, sodium salt (ANS). The critical micelle concentration was located at 0.069, 0.071 and 0.063 M by measurements of surface tension, perylene solubilization and enhancement of ANS fluorescence, respectively. In contrast to other cationic surfactants, the anesthetic monomer did not show evidence of forming a fluorescent molecular complex with ANS under the experimental conditions of this study. The formation of micelles by tetracaine-HCl showed a pronounced effect on lipid membranes by inducing an abrupt decrease in the scattered light of egg lecithin liposomes at an anesthetic concentration roughly similar to its critical micelle concentration. This optical behaviour is characteristic of liposome damage and can be interpreted to mean that the lipids become solubilized into tetracaine-HCl micelles. The ability of this local anesthetic to form micelles can be taken as a manifestation of the same hydrophobic forces that lead to partitioning of the drug into membranes.

Determination of surface potential in liposomes.

The ability of the fluorescent pH indicator 4-heptadecylumbelliferone to detect the electrical potential at the surface of negatively charged liposomes, was investigated. The vesicles were prepared from mixtures of egg lecithin and dicetyl phosphate at different molar ratios in NaCl solutions of various concentrations. It has been found that the dependence of the experimental surface potential in the proportion of charged lipid in the vesicles and on the salt concentration in the aqueous phase, was very similar to the predictions of the Gouy-Chapman equation as calculated by assuming a reasonable value for the mean molecular area of the lamellar lipids. In view of the good correlation obtained between the experimental and theoretical results, it is concluded that 43-heptadecylumbelliferone is quantitatively sensitive to changes in double-layer potential at the surface of lipid vesicles.

Disruption of liposomes by tetracaine micelles.

The effect of the local anesthetic tetracaine hydrochloride on the stability of egg lecithin vesicles has been studied by means of a filtration procedure using Pellicon membranes with an average molecular weight limit of retention of 10(5). These filters were efficient to retain liposomes and allowed the free passage of disrupted vesicles as detected by measuring the phospholipid content in the filtrates. It has been found that at concentrations above 0.05 M, the anesthetic induces disruption of liposomes presumably through formation of mixed micelles of tetracaine and phosphatidylcholine.

The effect of neutral and charged micelles on the acid-base dissociation of the local anesthetic tetracaine.

The influence of surfactant micelles on the acid-base dissociation of the charged tertiary amino group of the local anesthetic, tetracaine, has been investigated. From measurements of tetracaine fluorescence as a function of bulk pH, apparent pK values of 6.88, 7.58 and 9.92 were found in the presence of cationic, neutral and anionic micelles, respectively, in 10 mM NaCl. These values are considerably displaced with respect to the pK in aqueous solution which is 8.26. Such large shifts can be attributed to the effect of the surface polarity and electrical potential on the dissociation behavior of the anesthetic bound to micelles. It can be expected that the acid-base dissociation of a local anesthetic adsorbed to nerve fibers will also be affected by the properties of the membrane surface. Thus, it is suggested that the influence of the interfacial region on the pK of surface-bound molecules should not be disregarded when estimating the proportion of charged and uncharged forms of local anesthetics interacting with axonal membranes.

Activity of phospholipase A2 on a fluorescent substrate incorporated into non-hydrolyzable phospholipid liposomes.

The activity of phospholipase A2 (PLA2) on phospholipid liposomes depends on the physicochemical properties of the aggregated substrate, which are subject to continuous modification by the products released during hydrolysis. We propose here an experimental design that, by means of the incorporation of a fluorescent substrate at very low molar ratio (< or = 1:500) into a nonhydrolizable liposomal matrix of 1,2-dihexadecyl-sn-glycero-3-phosphocholine (DHPC), allows the study of hydrolysis by porcine pancreatic phospholipase A2, in virtual absence of physical perturbations of the lamellar phase, by the released products. We have been able to measure immediate hydrolysis of the fluorescent substrate 1,2-di-[omega(1'-pyreno)-decanoyl]-sn- glycero-3-phosphocholine when the sonicated liposomal matrix is in the gel phase. In the liquid crystalline state, in contrast, hydrolysis is very poor even after 80 min of adding the enzyme. Both in the gel and liquid-crystalline phases, incorporation of unlabeled PLA2 products activates the hydrolysis rate to comparable levels. It appears that the conformation adopted by the substrate immersed in the gel or liquid crystalline matrix is especially important in determining its susceptibility to hydrolysis in the absence of products.

The effect of pH on the phase transition temperature of dipalmitoylphosphatidylcholine-palmitic acid liposomes.

The shift in the gel-liquid crystal phase transition temperature (tm) of dipalmitoylphosphatidylcholine liposomes induced by incorporation of 10 mol% palmitic acid, was measured by 90 degrees light scattering at different bulk pH values. It has been found that the tm shift decreases sigmoidally from 4.7 to -0.3 degrees C as the bulk pH is raised from 5 to 11. Since it is in this range that the carboxyl group of a membrane-bound fatty acid should ionize, our results can be interpreted to mean that there is relationship between the tm shift and the degree of dissociation of palmitic acid, the uncharged fatty acid increasing tm and its conjugate, anionic form, slightly decreasing the transition temperature of dipalmitoylphosphatidylcholine liposomes. The experimental results are fitted by a modified form of the Henderson-Hasselbach equilibrium expression which takes into account the effect of the anionic fatty acid on the surface potential and hence, on the surface pH of liposomes, according to Gouy-Chapman and Boltzmann equations, respectively. Best fit between theory and experiments is found when the intrinsic interfacial pK of palmitic acid is set equal to 7.7. This high pK value can be explained as due to the effect of the lower dielectric constant of the interfacial region, as compared to bulk water, on the acid-base dissociation of the carboxyl group. The results presented here show that upon incorporation of palmitic acid, the phase transition of dipalmitoylphosphatidylcholine bilayers becomes extremely sensitive to changes of pH in the vicinity of the physiological range. This property is not shown by the pure phospholipid bilayers in the same pH range.

Membrane fluidity of Escherichia coli during heat-shock.

The excimer-forming fluorophore dipyrenylpropane has been used to measure the relative fluidity of total membranes isolated from Escherichia coli grown at 30 or 45 degrees C, or exposed to a heat-shock from 30 to 45 degrees C for various periods of time. Parallel experiments were performed using [35S]methionine pulse-labeling of cells, to study the induction of heat-shock proteins (HSPs) at different times after the sudden change in E. coli growth-temperature from 30 to 45 degrees C. Results suggest that upon an abrupt temperature upshift from 30 to 45 degrees C, membrane fluidity adjustment to the steady-state level at the high temperature, takes place during the E. coli heat-shock response.

Eggshells are shaped by a precise spatio-temporal arrangement of sequentially deposited macromolecules.

The avian eggshell is a composite bioceramic which is formed by a controlled interaction of an organic and an inorganic phase. The organic phase contains, among other constituents, type X collagen and proteoglycans, mainly keratan and dermatan sulfate. Understanding the principles governing the synthesis and temporo-spatial distribution of such macromolecules, and their influence on the organization of the crystalline phase, is an essential aspect of establishing the biological basis of the quality of eggshell, both as an embryonic chamber and as a natural food package. In the present study, we have examined the process of eggshell formation by immunohistochemistry, scanning electron microscopy and energy dispersive X-ray microanalysis. Precise sites and timing of secretion were established for the deposition of particular macromolecules. Type X collagen is detected at the very first moment of shell membrane formation. The appearance of keratan sulfate coincides with the appearance of mammillae, while dermatan sulfate is deposited later, coincident with shell matrix deposition. We propose that keratan sulfate, due to its precise localization, temporal appearance and calcium-binding affinity, relates to the maintenance of calcium reserve bodies, the primary source of calcium for the embryo. On the other hand, dermatan sulfate may control crystal growth, resulting in a preferential orientation of calcite crystals within the palisade layer.

Blastocoel formation in half and double mouse embryos.

We have tested the alleged causal relationship between the peripheral sealing of the embryo and the formation of the blastocoel by observing with the electron microscope the diffusion of a colloidal lanthanum tracer into advanced morulae and early blastocysts grown from half and double mouse embryos. In these experimental conditions and in the controls, the peripheral sealing of the embryos always precedes the appearance of the blastocoel, supporting a causal relationship between both phenomena. By counting the number of cells present at the morula-blastocyst transition in half and double embryos, as well as in control embryos, we have observed that neither the total number of cells nor the number of cell cycles correlate well with the appearance of the blastocoel. Results suggest that blastocoel formation depends on time elapsed since fertilization and that the experimental manipulation diminishes the cleavage rate.

The local anaesthetic tetracaine as a quencher of perylene fluorescence in micelles.

At neutral pH the local anaesthetic tetracaine hydrochloride quenches the fluorescence of the lipophilic dye perylene incorporated into non-ionic micelles. The process follows the Stern-Volmer equation, suggesting that quenching occurs through encounter of fluorophore and quencher. As the pH is lowered from 5 to 1, the apparent quenching constant decreases sigmoidally, the midpoint of the curve being at pH 2.3, close to the pK value characterizing the ionization of the anaesthetic aromatic butylamino group. Quenching is completely reversed below pH 1. These results show that the ability of tetracaine to quench the fluorescence of perylene incorporated into micelles depends on the absence of charge on its aromatic amine. Quenching was also studied in homogeneous dioxane-water solution. In this system the quenching constant also decreases sigmoidally as the pH is lowered. The infection point of the curve is nearly coincident with the pK of tetracaine butylamino group in the same partially non-aqueous medium. Protonation of this group induces 60% reversal of the quenching, suggesting that the main mechanism of fluorescence extinction could be the electron transfer from unprotonated tetracaine aromatic amine to perylene in the excited state. However, an additional process which remains operative even when such an amino group is positively charged must also be involved. It can be concluded that the complete reversal of tetracaine quenching of perylene fluorescence in micelles induced by low pH is due to the inability of the anaesthetic to become partitioned into micelles upon protonation of its aromatic amine. In contrast, at neutral pH the local anaesthetic is able to reach the micelle non-polar core where perylene is located. This is consistent with the models, suggesting that the membrane-bound tetracaine assumes a rod-like configuration parallel to the surface normal with the aromatic butylamino group located into a highly hydrophobic region.