Cell division protein FtsK coordinates bacterial chromosome segregation and daughter cell separation in Staphylococcus aureus.

Unregulated cell cycle progression may have lethal consequences and therefore, bacteria have various mechanisms in place for the precise spatiotemporal control of cell cycle events. We have uncovered a new link between chromosome replication/segregation and splitting of the division septum. We show that the DNA translocase domain-containing divisome protein FtsK regulates cellular levels of a peptidoglycan hydrolase Sle1, which is involved in cell separation in the bacterial pathogen Staphylococcus aureus. FtsK interacts with a chaperone (trigger factor, TF) and establishes a FtsK-dependent TF concentration gradient that is higher in the septal region. Trigger factor binds Sle1 and promotes its preferential export at the septal region, while also preventing Sle1 degradation by the ClpXP proteolytic machinery. Upon conditions that lead to paused septum synthesis, such as DNA damage or impaired DNA replication/segregation, TF gradient is dissipated and Sle1 levels are reduced, thus halting premature septum splitting.

Peptidoglycan synthesis drives an FtsZ-treadmilling-independent step of cytokinesis.

Peptidoglycan is the main component of the bacterial wall and protects cells from the mechanical stress that results from high intracellular turgor. Peptidoglycan biosynthesis is very similar in all bacteria; bacterial shapes are therefore mainly determined by the spatial and temporal regulation of peptidoglycan synthesis rather than by the chemical composition of peptidoglycan. The form of rod-shaped bacteria, such as Bacillus subtilis or Escherichia coli, is generated by the action of two peptidoglycan synthesis machineries that act at the septum and at the lateral wall in processes coordinated by the cytoskeletal proteins FtsZ and MreB, respectively. The tubulin homologue FtsZ is the first protein recruited to the division site, where it assembles in filaments-forming the Z ring-that undergo treadmilling and recruit later divisome proteins. The rate of treadmilling in B. subtilis controls the rates of both peptidoglycan synthesis and cell division. The actin homologue MreB forms discrete patches that move circumferentially around the cell in tracks perpendicular to the long axis of the cell, and organize the insertion of new cell wall during elongation. Cocci such as Staphylococcus aureus possess only one type of peptidoglycan synthesis machinery, which is diverted from the cell periphery to the septum in preparation for division. The molecular cue that coordinates this transition has remained elusive. Here we investigate the localization of S. aureus peptidoglycan biosynthesis proteins and show that the recruitment of the putative lipid II flippase MurJ to the septum, by the DivIB-DivIC-FtsL complex, drives peptidoglycan incorporation to the midcell. MurJ recruitment corresponds to a turning point in cytokinesis, which is slow and dependent on FtsZ treadmilling before MurJ arrival but becomes faster and independent of FtsZ treadmilling after peptidoglycan synthesis activity is directed to the septum, where it provides additional force for cell envelope constriction.

Staphylococcus aureus requires at least one FtsK/SpoIIIE protein for correct chromosome segregation.

Faithful coordination between bacterial cell division and chromosome segregation in rod-shaped bacteria, such as Escherichia coli and Bacillus subtilis, is dependent on the DNA translocase activity of FtsK/SpoIIIE proteins, which move DNA away from the division site before cytokinesis is completed. However, the role of these proteins in chromosome partitioning has not been well studied in spherical bacteria. Here, it was shown that the two Staphylococcus aureus FtsK/SpoIIIE homologues, SpoIIIE and FtsK, operate in independent pathways to ensure correct chromosome management during cell division. SpoIIIE forms foci at the centre of the closing septum in at least 50% of the cells that are close to complete septum synthesis. FtsK is a multifunctional septal protein with a C-terminal DNA translocase domain that is not required for correct chromosome management in the presence of SpoIIIE. However, lack of both SpoIIIE and FtsK causes severe nucleoid segregation and morphological defects, showing that the two proteins have partially redundant roles in S. aureus.

Sequential swallowing of liquid in elderly adults: cup or straw?

This article describes the study of the characteristics of sequential swallowing of 100 ml of liquid (dyed water) in two swallowing trials, directly from a cup and through a straw, in healthy elderly individuals. The aim of the study was to determine whether differences in the swallowing pattern are influenced by the type of utensil used. The subjects were subjected to clinical assessment and fiberoptic endoscopic evaluation of swallowing. The research found that intake from the cup showed a significantly lower median as regards time to drink the total volume. The final intake volume was significantly larger from the cup. A statistically significant difference was found in the oral spillage of liquid, which was notably higher in the cup trial. Despite the presence of residue in the valleculae and pyriform sinuses, in neither trial was there penetration or aspiration of liquid. The straw has a favorable influence on the quality of the sequential swallowing of liquid in regard to bolus containment within the oral cavity, which was better with that utensil. The cup provides a higher final volume in a shorter time for intake but there is more fluid spillage.

A CRISPRi-based genetic resource to study essential Staphylococcus aureus genes.

IMPORTANCE: Staphylococcus aureus is an important clinical pathogen that causes a high number of antibiotic-resistant infections. The study of S. aureus biology, and particularly of the function of essential proteins, is of particular importance to develop new approaches to combat this pathogen. We have optimized a clustered regularly interspaced short palindromic repeat interference (CRISPRi) system that allows efficient targeting of essential S. aureus genes. Furthermore, we have used that system to construct a library comprising 261 strains, which allows the depletion of essential proteins encoded by 200 genes/operons. This library, which we have named Lisbon CRISPRi Mutant Library, should facilitate the study of S. aureus pathogenesis and biology.

The role of GpsB in Staphylococcus aureus cell morphogenesis.

For decades, cells of the Gram-positive bacterial pathogen Staphylococcus aureus were thought to lack a dedicated elongation machinery. However, S. aureus cells were recently shown to elongate before division, in a process that requires a shape elongation division and sporulation (SEDS)/penicillin-binding protein (PBP) pair for peptidoglycan synthesis, consisting of the glycosyltransferase RodA and the transpeptidase PBP3. In ovococci and rod-shaped bacteria, the elongation machinery, or elongasome, is composed of various proteins besides a dedicated SEDS/PBP pair. To identify proteins required for S. aureus elongation, we screened the Nebraska Transposon Mutant Library, which contains transposon mutants in virtually all non-essential staphylococcal genes, for mutants with modified cell shape. We confirmed the roles of RodA/PBP3 in S. aureus elongation and identified GpsB, SsaA, and RodZ as additional proteins involved in this process. The gpsB mutant showed the strongest phenotype, mediated by the partial delocalization from the division septum of PBP2 and PBP4, two penicillin-binding proteins that synthesize and cross-link peptidoglycan. Increased levels of these PBPs at the cell periphery versus the septum result in higher levels of peptidoglycan insertion/crosslinking throughout the entire cell, possibly overriding the RodA/PBP3-mediated peptidoglycan synthesis at the outer edge of the septum and/or increasing stiffness of the peripheral wall, impairing elongation. Consequently, in the absence of GpsB, S. aureus cells become more spherical. We propose that GpsB has a role in the spatio-temporal regulation of PBP2 and PBP4 at the septum versus cell periphery, contributing to the maintenance of the correct cell morphology in S. aureus. IMPORTANCE: Staphylococcus aureus is a Gram-positive clinical pathogen, which is currently the second cause of death by antibiotic-resistant infections worldwide. For decades, S. aureus cells were thought to be spherical and lack the ability to undergo elongation. However, super-resolution microscopy techniques allowed us to observe the minor morphological changes that occur during the cell cycle of this pathogen, including cell elongation. S. aureus elongation is not required for normal growth in laboratory conditions. However, it seems to be essential in the context of some infections, such as osteomyelitis, during which S. aureus cells apparently elongate to invade small channels in the bones. In this work, we uncovered new determinants required for S. aureus cell elongation. In particular, we show that GpsB has an important role in the spatio-temporal regulation of PBP2 and PBP4, two proteins involved in peptidoglycan synthesis, contributing to the maintenance of the correct cell morphology in S. aureus.

The Holliday junction resolvase RecU is required for chromosome segregation and DNA damage repair in Staphylococcus aureus.

BACKGROUND: The Staphylococcus aureus RecU protein is homologous to a Bacillus subtilis Holliday junction resolvase. Interestingly, RecU is encoded in the same operon as PBP2, a penicillin-binding protein required for cell wall synthesis and essential for the full expression of resistance in Methicillin Resistant S. aureus strains. In this work we have studied the role of RecU in the clinical pathogen S. aureus. RESULTS: Depletion of RecU in S. aureus results in the appearance of cells with compact nucleoids, septa formed over the DNA and anucleate cells. RecU-depleted cells also show increased septal recruitment of the DNA translocase SpoIIIE, presumably to resolve chromosome segregation defects. Additionally cells are more sensitive to DNA damaging agents such as mitomycin C or UV radiation. Expression of RecU from the ectopic chromosomal spa locus showed that co-expression of RecU and PBP2 was not necessary to ensure correct cell division, a process that requires tight coordination between chromosome segregation and septal cell wall synthesis. CONCLUSIONS: RecU is required for correct chromosome segregation and DNA damage repair in S. aureus. Co-expression of recU and pbp2 from the same operon is not required for normal cell division.

Teichoic acids are temporal and spatial regulators of peptidoglycan cross-linking in Staphylococcus aureus.

The cell wall of Staphylococcus aureus is characterized by an extremely high degree of cross-linking within its peptidoglycan (PGN). Penicillin-binding protein 4 (PBP4) is required for the synthesis of this highly cross-linked peptidoglycan. We found that wall teichoic acids, glycopolymers attached to the peptidoglycan and important for virulence in Gram-positive bacteria, act as temporal and spatial regulators of PGN metabolism, controlling the level of cross-linking by regulating PBP4 localization. PBP4 normally localizes at the division septum, but in the absence of wall teichoic acids synthesis, it becomes dispersed throughout the entire cell membrane and is unable to function normally. As a consequence, the peptidoglycan of TagO null mutants, impaired in wall teichoic acid biosynthesis, has a decreased degree of cross-linking, which renders it more susceptible to the action of lysozyme, an enzyme produced by different host organisms as an initial defense against bacterial infection.

Modelling the effect of defects and cracks in solar cells' performance using the d1MxP discrete model.

Renewable energies are increasingly playing an important role in the world's energy supply. Society demands new solutions to solve environmental issues caused by fossil fuels. The importance of photovoltaic technology has been increasing and consequently, the necessity to have more accurate models to characterise the performance of solar cells during their entire lifetime has rose as well. Performance problems may appear during devices' lifetimes associated with factors, such as weather conditions or faulty installation. Cracking might occur, leading to abrupt reductions on the produced power, quite difficult and expensive to fix. The I-V curves of a defected or cracked solar cell might not have the shape imposed by the usual models as 1M5P. In this article, cracked c-Si solar cells are modelled using a novel model: d1MxP. This model is based on the discretisation of the diode's response on models as 1M5P. Instead of imposing a shape and compute some parameters to fit it on experimental data, the proposed model connects every two points. The results suggest a better fit using the proposed model in comparison with the 1M5P, not only in the original curves, but also modelling cracked cells. As consequence of a better fitting, the computation of important figures of merit as maximum power point or fill factor, reveals to be more precise. It is concluded that the proposed model might characterise the performance of a solar cell, even cracked, which is a huge advance aiming the possibility of simulating complex problems during the cells' operation lifetime.

Absence of nucleoid occlusion effector Noc impairs formation of orthogonal FtsZ rings during Staphylococcus aureus cell division.

The Gram-positive pathogen Staphylococcus aureus divides by synthesizing the septum in three orthogonal planes over three consecutive division cycles. This process has to be tightly coordinated with chromosome segregation to avoid bisection of the nucleoid by the septum. Here we show that deletion of the nucleoid occlusion effector Noc in S. aureus results in the formation of Z-rings over the nucleoid, as well as in DNA breaks, indicating that Noc has an important role as an antiguillotine checkpoint that prevents septa from forming over the DNA. Furthermore, Noc deleted cells show multiple Z-rings which are no longer placed in perpendicular planes. We propose that the axis of chromosome segregation has a role in determining the placement of the division septum. This is achieved via the action of Noc which restricts the placement of the division septum to one of an infinite number of potential division planes that exist in S. aureus.

Synthetic antimicrobial peptides as enhancers of the bacteriolytic action of staphylococcal phage endolysins.

Bacteriophage endolysins degrade the bacterial cell wall and are therefore considered promising antimicrobial alternatives to fight pathogens resistant to conventional antibiotics. Gram-positive bacteria are usually considered easy targets to exogenously added endolysins, since their cell walls are not shielded by an outer membrane. However, in nutrient rich environments these bacteria can also tolerate endolysin attack if they keep an energized cytoplasmic membrane. Hence, we have hypothesized that the membrane depolarizing action of antimicrobial peptides (AMPs), another attractive class of alternative antibacterials, could be explored to overcome bacterial tolerance to endolysins and consequently improve their antibacterial potential. Accordingly, we show that under conditions supporting bacterial growth, Staphylococcus aureus becomes much more susceptible to the bacteriolytic action of endolysins if an AMP is also present. The bactericidal gain resulting from the AMP/endolysin combined action ranged from 1 to 3 logs for different S. aureus strains, which included drug-resistant clinical isolates. In presence of an AMP, as with a reduced content of cell wall teichoic acids, higher endolysin binding to cells is observed. However, our results indicate that this higher endolysin binding alone does not fully explain the higher susceptibility of S. aureus to lysis in these conditions. Other factors possibly contributing to the increased endolysin susceptibility in presence of an AMP are discussed.

Monofunctional transglycosylases are not essential for Staphylococcus aureus cell wall synthesis.

The polymerization of peptidoglycan is the result of two types of enzymatic activities: transglycosylation, the formation of linear glycan chains, and transpeptidation, the formation of peptide cross-bridges between the glycan strands. Staphylococcus aureus has four penicillin binding proteins (PBP1 to PBP4) with transpeptidation activity, one of which, PBP2, is a bifunctional enzyme that is also capable of catalyzing transglycosylation reactions. Additionally, two monofunctional transglycosylases have been reported in S. aureus: MGT, which has been shown to have in vitro transglycosylase activity, and a second putative transglycosylase, SgtA, identified only by sequence analysis. We have now shown that purified SgtA has in vitro transglycosylase activity and that both MGT and SgtA are not essential in S. aureus. However, in the absence of PBP2 transglycosylase activity, MGT but not SgtA becomes essential for cell viability. This indicates that S. aureus cells require one transglycosylase for survival, either PBP2 or MGT, both of which can act as the sole synthetic transglycosylase for cell wall synthesis. We have also shown that both MGT and SgtA interact with PBP2 and other enzymes involved in cell wall synthesis in a bacterial two-hybrid assay, suggesting that these enzymes may work in collaboration as part of a larger, as-yet-uncharacterized cell wall-synthetic complex.

Fluorescent reporters for studies of cellular localization of proteins in Staphylococcus aureus.

We have constructed a set of plasmids that allow expression, from their native chromosomal loci, of Staphylococcus aureus proteins fused to one of four different fluorescent proteins (green fluorescent protein [GFP], cyan fluorescent protein [CFP], yellow fluorescent protein [YFP], and mCherry), using two different resistance markers (kanamycin and erythromycin). We have also constructed a plasmid that allows expression of proteins from the ectopic spa locus in the S. aureus chromosome. This toolbox can be used for studies of the localization of proteins in S. aureus, a prominent pathogen in both health care and community settings.

Inactivation of the SauI type I restriction-modification system is not sufficient to generate Staphylococcus aureus strains capable of efficiently accepting foreign DNA.

Genetic manipulation of Staphylococcus aureus is limited by the availability of only a single strain, RN4220, that is capable of easily accepting foreign DNA. Inactivation of the hsdR gene of the SauI type I restriction-modification system was shown previously to be responsible for the high transformation efficiency of RN4220 (D. E. Waldron and J. A. Lindsay, J Bacteriol. 188:5578-5585, 2006). However, deletion of this gene in three different S. aureus strains was not sufficient to make them readily transformable, which would be remarkably useful for genetic studies of this pathogenic organism. These results indicate that another unknown factor(s) is required for the transformable phenotype in S. aureus.

Cell shape dynamics during the staphylococcal cell cycle.

Staphylococcus aureus is an aggressive pathogen and a model organism to study cell division in sequential orthogonal planes in spherical bacteria. However, the small size of staphylococcal cells has impaired analysis of changes in morphology during the cell cycle. Here we use super-resolution microscopy and determine that S. aureus cells are not spherical throughout the cell cycle, but elongate during specific time windows, through peptidoglycan synthesis and remodelling. Both peptidoglycan hydrolysis and turgor pressure are required during division for reshaping the flat division septum into a curved surface. In this process, the septum generates less than one hemisphere of each daughter cell, a trait we show is common to other cocci. Therefore, cell surface scars of previous divisions do not divide the cells in quadrants, generating asymmetry in the daughter cells. Our results introduce a need to reassess the models for division plane selection in cocci.

Characteristics of sequential swallowing of liquids in young and elderly adults: an integrative review.

PURPOSE: To perform an integrative review of studies on liquid sequential swallowing, by characterizing the methodology of the studies and the most important findings in young and elderly adults. RESEARCH STRATEGY: Review of the literature written in English and Portuguese on PubMed, LILACS, SciELO and MEDLINE databases, within the past twenty years, available fully, using the following uniterms: sequential swallowing, swallowing, dysphagia, cup, straw, in various combinations. SELECTION CRITERIA: Research articles with a methodological approach on the characterization of liquid sequential swallowing by young and/or elderly adults, regardless of health condition, excluding studies involving only the esophageal phase. DATA ANALYSIS: The following research indicators were applied: objectives, number and gender of participants; age group; amount of liquid offered; intake instruction; utensil used, methods and main findings. RESULTS: 18 studies met the established criteria. The articles were categorized according to the sample characterization and the methodology on volume intake, utensil used and types of exams. Most studies investigated only healthy individuals, with no swallowing complaints. Subjects were given different instructions as to the intake of all the volume: usual manner, continually, as rapidly as possible. The findings about the characterization of sequential swallowing were varied and described in accordance with the objectives of each study. CONCLUSION: It found great variability in the methodology employed to characterize the sequential swallowing. Some findings are not comparable, and sequential swallowing is not studied in most swallowing protocols, without consensus on the influence of the utensil.

Phosphonium-based ionic liquids as modifiers for biomedical grade poly(vinyl chloride).

This work reports and discusses the influence of four phosphonium-based ionic liquids (PhILs), namely trihexyl(tetradecyl) phosphonium dicyanamide, [P(6,6,6,14)][dca]; trihexyl(tetradecyl) phosphonium bis(trifluoromethylsulfonyl)imide, [P(6,6,6,14)][Tf(2)N]; tetrabutyl phosphonium bromide, [P(4,4,4,4)][Br]; and tetrabutyl phosphonium chloride, [P(4,4,4,4)][Cl], on some of the chemical, physical and biological properties of a biomedical-grade suspension of poly(vinyl chloride) (PVC). The main goal of this work was to evaluate the capacity of these PhILs to modify some of the properties of neat PVC, in particular those that may allow their use as potential alternatives to traditional phthalate-based plasticizers in PVC biomedical applications. PVC films having different PhIL compositions (0, 5, 10 and 20 wt.%) were prepared (by solvent film casting) and characterised by Fourier transform infrared, thermogravimetric analysis, differential scanning calorimetry, dynamical mechanical thermal analysis, scanning electron microscopy/energy-dispersive X-ray/electron probe microanalysis, X-ray diffraction, transmittance, permeability towards oxygen and carbon dioxide, thermal degradation, contact angle measurement, water and vapour uptake, leachability and biocompatibility (haemolytic potential, thrombogenicity and cytotoxicity). A conventional organic plasticizer (di-isononyl phthalate) was used for comparison purposes. The results obtained showed that it was possible to change the neat PVC hydrophobicity, and consequently its water uptake capacity and plasticizer leachability, just by changing the PhIL employed and its composition. It was also possible to significantly change the thermal and mechanical properties of PVC films by choosing appropriate PhIL cation/anion combinations. However, a specific PhIL may not always be capable of simultaneously keeping and/or improving both physical properties. In addition, ionic halide salts were found to promote PVC dehydrochlorination. Finally, none of the prepared materials presented toxicity against Caco-2 cells, though pure [P(6,6,6,14)][dca] decreased HepG2 cells viability. Moreover, PVC films with [P(6,6,6,14)][dca] and [P(4,4,4,4)][Cl] were found to be haemolytic and thus these PhILs must be avoided as PVC modifiers if biomedical applications are envisaged. In conclusion, from all the PhILs tested, [P(6,6,6,14)][Tf(2)N] showed the most promising results regarding blood compatibility, leaching and permeability to gases of PVC films. The results presented are a strong indicator that adequate PhILs may be successfully employed as PVC multi-functional plasticizers for a wide range of potential applications, including those in the biomedical field.

Dynamical characterization of a cellulose acetate polysaccharide.

This work brings together dynamical and structural information at a molecular level for cellulose acetate being an original contribution to the general description of polysaccharide properties. In particular, it allowed reinterpreting the secondary relaxation mechanisms that are still controversial in the literature; a compilation of data provided by different authors is provided. Detailed dynamical information is provided by dielectric relaxation spectroscopy (DRS) (10(-1)-10(6) Hz) for cellulose acetate (CA) in the sub-T(g) region below ambient temperature; results were compared with cellulose acetate structured as an asymmetric membrane (CAmb). In samples with low water content, two secondary relaxation processes between 173 and 298 K were identified by DRS, associated with localized mobility. The process located at the lowest temperatures (process I) has a different mobility in CA relative to CAmb. The identical crystalline/amorphous state of both materials allowed rationalizing the distinct behavior in terms of polymeric arrangement and ability for water uptake. The looser structure of the CA relative to CAmb as confirmed by FTIR, TGA, and DSC analysis makes more sites accessible to water molecules, resulting in a higher water retention in CA (2.73% w/w) relative to CAmb (1.60% w/w) and an increased molecular mobility in the former due to a plasticizing effect. In both materials, process I is significantly influenced by hydration, shifting to higher frequencies and lower temperatures upon water uptake. This process seems to be associated with mobility occurring within the monomeric unit, which embraces the two anhydroglucose rings connected by the glycosidic linkage and the polar groups directly attached to it. It should involve a very limited length scale, as suggested by its location, far below the glass transition, and the tau(infinity) value with a low entropic effect. The relaxation process that emerges later, process II, is similar for both samples being much less influenced by water but experiencing a slight antiplasticizing effect shifting to lower frequencies and higher temperatures upon hydration. It should involve side group motions, strongly coupled to the mobility of the anhydroglucose rings, which become hindered probably due to establishment of H-bonds with water molecules. The plasticizing/antiplasticizing effect is being discussed only on the basis of the frequency position of the relaxation peak. Processes I and II merge into a broad relaxation (gamma(dry)) upon water removal in both CA and CAmb, however evolving slower in the former with drying, due to a more disordered structure of CA that allows water to interact with more internal sites in the polymer. At higher temperatures (T > or = 353 K), a process emerges in the high frequency side of the dynamic alpha-relaxation which is compatible with a beta(JG)-relaxation. The structured specimen CAmb provided an additional way to probe the morphological changes undergone by the material when annealed to temperatures higher than 353 K, originating an increase in the dielectric response. This effect can be associated with a skin densification and partial collapse of the membrane porous network, as observed by SEM.

Características da deglutição sequencial de líquidos em adultos e idosos: uma revisão integrativa / Characteristics of sequential swallowing of liquids in young and elderly adults: an integrative review

OBJETIVO: Realizar revisão integrativa de estudos sobre deglutição sequencial de líquidos, caracterizando o método e os principais achados em adultos e idosos. ESTRATÉGIA DE PESQUISA: Foi realizado levantamento na literatura internacional, publicada nos idiomas Inglês ou Português, utilizando-se base de dados PubMed, LILACS, SciELO, MEDLINE, dos últimos 20 anos, cujos artigos estavam disponíveis na íntegra. Foram utilizadas as seguintes palavras, em combinações variadas: deglutição sequencial, deglutição, disfagia, copo e canudo. CRITÉRIOS DE SELEÇÃO: Foram incluídos artigos de pesquisa com abordagem metodológica referente à caracterização da deglutição sequencial de líquidos em sujeitos adultos e/ou idosos, independentemente de patologia envolvida. Excluíram-se os estudos que englobavam apenas a fase esofágica. ANÁLISE DOS DADOS: Foram considerados indicadores da pesquisa: objetivos, número e gênero dos participantes; faixa etária; quantidade de líquido ofertado; instrução para ingesta; utensílio e métodos utilizados; principais resultados. RESULTADOS: Dezoito estudos contemplaram os critérios propostos. Os artigos foram tabulados quanto à caracterização da amostra e à metodologia (volume ofertado, utensílio utilizado e tipos de exames). A maioria dos artigos envolveu apenas indivíduos saudáveis e sem queixas de deglutição. Houve diferentes tipos de instrução para deglutir todo o volume: maneira habitual; continuamente; o mais rápido possível. Os achados referentes às caracterizações da deglutição sequencial foram variados e descritos de acordo com objetivos de cada estudo. CONCLUSÃO: Constata-se grande variabilidade na metodologia empregada para caracterização da deglutição sequencial. Alguns achados não são comparáveis entre si, sendo que a deglutição sequencial ainda não faz parte da maioria dos protocolos de avaliação da deglutição. Não há consenso sobre a influência do utensílio.

PURPOSE: To perform an integrative review of studies on liquid sequential swallowing, by characterizing the methodology of the studies and the most important findings in young and elderly adults. RESEARCH STRATEGY: Review of the literature written in English and Portuguese on PubMed, LILACS, SciELO and MEDLINE databases, within the past twenty years, available fully, using the following uniterms: sequential swallowing, swallowing, dysphagia, cup, straw, in various combinations. SELECTION CRITERIA: Research articles with a methodological approach on the characterization of liquid sequential swallowing by young and/or elderly adults, regardless of health condition, excluding studies involving only the esophageal phase. DATA ANALYSIS: The following research indicators were applied: objectives, number and gender of participants; age group; amount of liquid offered; intake instruction; utensil used, methods and main findings. RESULTS: 18 studies met the established criteria. The articles were categorized according to the sample characterization and the methodology on volume intake, utensil used and types of exams. Most studies investigated only healthy individuals, with no swallowing complaints. Subjects were given different instructions as to the intake of all the volume: usual manner, continually, as rapidly as possible. The findings about the characterization of sequential swallowing were varied and described in accordance with the objectives of each study. CONCLUSION: It found great variability in the methodology employed to characterize the sequential swallowing. Some findings are not comparable, and sequential swallowing is not studied in most swallowing protocols, without consensus on the influence of the utensil.