Application of robotic surgery and traditional laparoscopic surgery in lymph node dissection for gynecological cancer: A meta­analysis.

Since the advantages of robotic surgery and laparoscopic surgery in the number of lymph node resections are not well understood, this meta-analysis used evidence-based medicine to assess the difference in the number of lymph nodes retrieved in gynecological cancer between the two surgical methods to guide clinical treatment. In the present meta-analysis, the Pubmed, Embase, Cochrane, China National Knowledge Infrastructure and Wanfang libraries were searched for articles that were published from the time of the database's inception to January 2021, including cohort studies and randomized controlled trials, where the observation group underwent robotic surgery to treat gynecological cancers and the control group underwent laparoscopic surgery to treat gynecological cancers, including cervical and ovarian cancers and endometrial cancers. Duplicate publications, studies with no full text, incomplete information or where the authors were unable to perform data extraction, animal experiments, reviews and systematic reviews were excluded. STATA 15.1 was used to analyze the data. Robotic surgery resulted in a significant increase in the number of lymph nodes retrieved from the pelvis [standard mean difference (SMD)=0.24; 95% CI, 0.04-0.45; P=0.007] and para-aortic (SMD=0.41; 95% CI, 0.13-0.69; P=0.004) regions compared with the number retrieved by laparoscopic surgery. Furthermore, there was no significant difference in operating time between robotic and laparoscopic surgery, despite the use of different instruments (SMD=0.12; 95% CI, -0.35-0.58; P=0.616). The amount of blood lost during robotic surgery was significantly less compared with that lost during laparoscopic surgery [SMD=-0.40; 95% CI, -0.58-(-0.22); P<0.001]. The present study evaluated cancer recurrence and death in further detail, and no statistically significant difference was demonstrated between robotic surgery and laparoscopic surgery in terms of recurrence rate [odds ratio (OR)=0.59; 95% CI, 0.21-1.65; P=0.318] and mortality rate (OR=0.31; 95% CI, 0.08-1.30; P=0.109). The present study demonstrated that robotic surgery was able to retrieve more pelvic and para-aortic lymph nodes than traditional laparoscopic surgery, which was consistent with previous reports. With regards to blood loss, The difference in operation time between the two surgical methods was not statistically significant, whereas the estimated blood loss of robotic surgery was significantly lower than that of traditional laparoscopic surgery. There was no statistically significant difference in the recurrence rate and mortality rate of the two surgical modality.

Assessing the bioavailability of antibiotics in soil with the diffusive gradients in thin films (DGT).

The diffusive gradients in thin films (DGT) technique is an excellent method for investigating the dynamic processes of antibiotics in soils. However, whether it is applicable in antibiotic bioavailability assessment is yet to be disclosed. This study employed DGT to determine the antibiotic bioavailability in soil, and compared the results with plant uptake, soil solutions, and solvent extraction methods. DGT exhibited predictive capability for plant taking in antibiotics proved by the significant linear relationship between the DGT based concentration (CDGT) and antibiotic concentration in roots and shoots. Although the performance of soil solution was acceptable based on linear relationship analysis, its stability was weaker than DGT. The results based on plant uptake and DGT indicated the bioavailable antibiotic contents in different soils were inconsistent because of the distinct mobility and resupply of sulphonamides and trimethoprim in different soils, as represented by Kd and Rds, which were affected by soil properties. Plant species played an important role in antibiotic uptake and translocation. Antibiotic uptake by plants depends on antibiotic, plant and soil. These results confirmed the capability of DGT in determining antibiotic bioavailability for the first time. This work provided a simple and powerful tool for environmental risk evaluation of antibiotics in soils.

Comparative transcriptomic analysis of two Cucumis melo var. saccharinus germplasms differing in fruit physical and chemical characteristics.

BACKGROUND: Hami melon (Cucumis melo var. saccharinus) is a popular fruit in China because of its excellent taste, which is largely determined by its physicochemical characteristics, including flesh texture, sugar content, aroma, and nutrient composition. However, the mechanisms by which these characteristics are regulated have not yet been determined. In this study, we monitored changes in the fruits of two germplasms that differed in physicochemical characteristics throughout the fruit development period. RESULTS: Ripe fruit of the bred variety 'Guimi' had significantly higher soluble sugar contents than the fruit of the common variety 'Yaolong.' Additionally, differences in fruit shape and color between these two germplasms were observed during development. Comparative transcriptome analysis, conducted to identify regulators and pathways underlying the observed differences at corresponding stages of development, revealed a higher number of differentially expressed genes (DEGs) in Guimi than in Yaolong. Moreover, most DEGs detected during early fruit development in Guimi were associated with cell wall biogenesis. Temporal analysis of the identified DEGs revealed similar trends in the enrichment of downregulated genes in both germplasms, although there were differences in the enrichment trends of upregulated genes. Further analyses revealed trends in differential changes in multiple genes involved in cell wall biogenesis and sugar metabolism during fruit ripening. CONCLUSIONS: We identified several genes associated with the ripening of Hami melons, which will provide novel insights into the molecular mechanisms underlying the development of fruit characteristics in these melons.

Neutrophil-derived heparin binding protein triggers vascular leakage and synergizes with myeloperoxidase at the early stage of severe burns (With video).

BACKGROUND: Burn shock caused by vascular leakage is one of the main causes of high mortality in severe burn injury. However, the pathophysiological mechanism of vascular leakage is still unclear. The purpose of this study was to explore the molecular mechanism of vascular leakage in the early stage of severe burn and provide a new target for the treatment of severe burns. METHODS: Neutrophils were isolated from human peripheral blood by magnetic beads sorting. ELISA was used to detect neutrophil-derived granule proteins and glycocalyx injury products in plasma. The vascular leakage and neutrophil movement were assessed by in vivo laser confocal imaging in mice, and high-quality video were provided.. Adhesion-related molecules were investigated by qRT-PCR. The damage to glycocalyx of mice vascular endothelial cells was observed by transmission electron microscope and scanning electron microscope. Proteomic analysis, flow cytometry and immunofluorescence were used to further study the relationship between human peripheral blood neutrophil-derived hypochlorite (HOCl) and CD44 of human vascular endothelial cells. RESULTS: In this study, we found that rapidly increasing activated neutrophils secrete heparin binding protein (HBP) and myeloperoxidase (MPO) after severe burn injury. Increased HBP triggers vascular leakage with synergy of MPO, results in systemic edema and burn shock. Furthermore, we found that the MPO catalytic product HOCl but not MPO triggers CD44 extracellular domain shedding from vascular endothelial cells to damage the glycocalyx. Damage to the glycocalyx results in firm adhesion of neutrophils and increases vascular leakage. However, MPO inhibitors partially protect the glycocalyx of vascular endothelial cells. The combination of HBP and MPO inhibitors markedly reduces vascular leakage and systemic edema in the early stage of severe burns. CONCLUSIONS: Taken together, these data reveal that neutrophil-derived HBP and MPO play an important synergies role in triggering vascular leakage at the early stage of severe burns. Targeted intervention in these two biomolecules may introduce new strategies for helping to reduce large amount of fluid loss and subsequent burn shock.

Amphipathic Janus Membrane with Hierarchical Multiscale Hyperporous Structure for Interfacial Catalysis.

The rational design and realization of multiscale porous structures has been a long-standing challenge in membrane science. Block copolymers (BCPs) with their self-assembly-enabled nanodomains have the potential to make structural breakthroughs. An amphipathic Janus membrane, with a hierarchical multiscale hyperporous structure constituted by polystyrene-b-poly(4-vinylpyridine) (PS4VP) and polyvinylidene fluoride (PVDF) blocks, was designed and synthesized in this work. Hydrophobic PVDF dominated one side of the membrane, and hydrophilic PS4VP, with nanopores that formed inside the macroporous channels of PVDF via a self-assembly approach, dominated the other side. Candida Rugosa Lipase (CRL), as a model biocatalyst, was immobilized in the PS4VP nanopores via injection. The immobilized lipase was exactly suspended at the interface of the organic and aqueous phases, owing to the amphipathic property of the Janus membrane. The designed structures and catalysis performances were further characterized. The immobilized lipase exhibited a three times higher specific activity than free lipase, and the relative activity still remained above 90% after 10 cycles of reusing, indicating the observable promotion and the guaranteed stability of the Janus membrane in interfacial catalysis. This work provided a general, facile and unique example for the design and synthesis of a hierarchical multiscale hyperporous membrane for interfacial catalysis.

Deep Neck Infection in Systemic Lupus Erythematosus Patients: Real-World Evidence.

Systemic lupus erythematosus (SLE) might increase deep neck infection (DNI) risk, but evidence supporting this hypothesis is limited. In this retrospective follow-up study, the SLE-DNI association was investigated using data from the Registry for Catastrophic Illness Patients, which is a subset of the Taiwan National Health Insurance Research Database. All patients newly diagnosed as having SLE in 1997-2011 were identified, and every SLE patient was individually matched to four patients without SLE according to sex, age, and socioeconomic status. The study outcome was DNI occurrence. DNI treatment modalities and prognoses in SLE and non-SLE patients, along with the association of steroid dose with DNI risk, were also studied. In total, 17,426 SLE and 69,704 non-SLE patients were enrolled. Cumulative DNI incidence was significantly higher in the SLE cohort than in the non-SLE cohort (p < 0.001). The Cox regression model demonstrated that SLE significantly increased DNI risk (hazard ratio: 4.70; 95% confidence interval: 3.50-6.32, p < 0.001). Moreover, in the sensitivity and subgroup analyses, the effect of SLE on DNI was stable. Relatively few SLE-DNI patients received surgical interventions (15.6% vs. 28.6%, p = 0.033). The between-group differences in tracheostomy use and hospitalisation duration were nonsignificant. In SLE patients, high steroid doses significantly increased DNI incidence (≥3 vs. <3 mg/day = 2.21% vs. 0.52%, p < 0.001). This is the first study demonstrating that SLE increases DNI risk by approximately five times and that high steroid dose increases DNI incidence in SLE patients.

Efficacy of Vibrio parahaemolyticus depuration in oysters (Crassostrea gigas).

This study investigated the influences of seawater to oyster ratio on depuration for decontaminating V. parahaemolyticus in raw oysters with a goal of identifying the proper ratio of oyster to seawater capable of improving the efficacy of the depuration process. The water to oyster ratios tested in this study ranged from 1.0 to 2.5 L of artificial seawater (ASW) per oyster (40 oysters in 40, 60, 80 and 100 L ASW). The depuration efficacy for purging V. parahaemolyticus from oysters was highest when we applied a 2:1, followed by 1.5:1, 2.5:1, and 1:1 L of ASW/oyster. Further studies of depuration with 2:1 L of ASW/oyster found that the concentration of V. parahaemolyticus in oysters decreased in a nonlinear manner. The depuration curve was fitted to a one phase decay model with a coefficient of determination (R2) of 0.933. The time for a 3 log reduction was 1.75 days with a 95% confidence interval from 1.65 to 1.85 days, which meets the FDA's requirement of larger than a 3.0 log (MPN/g) reduction as a post-harvest process for V. parahaemolyticus control. After 4 days levels in all trials were <100 MPN/g meeting performance standards established by Japan and Canada. Furthermore, the time for a 3.52 log reduction was 3.17 days with a 95% confidence interval from 2.92 to 3.54 days but it took 5 days to reduce levels to <30 MPN/g, which satisfies FDA's requirement as a post-harvest control process (>3.52 log MPN/g reduction) for the purpose of making safety added labeling claims for V. parahaemolyticus.

Metal-Organic Framework Nanocomposite Thin Films with Interfacial Bindings and Self-Standing Robustness for High Water Flux and Enhanced Ion Selectivity.

Metal-organic framework (MOF)-based materials are promising candidates for a range of separation applications. However, the fabrication of self-standing MOF-based thin films remains challenging. Herein, a facile solution casting strategy is developed for fabricating UiO-66 nanocomposite thin films (UiO66TFs) with thicknesses down to ∼400 nm. Nanosizing UiO-66 and incorporating sulfonated polysulfone additives render high dispersity and interfacial bindings between MOFs and polymer matrices, so UiO66TFs are more mechanically robust and thermally stable than their pure-polymer counterparts. Enhanced microporosity with sub-nanometer pore sizes of the self-standing membranes enables the direct translation of UiO-66-based sorption and ion-sieving properties, thus increasing water flux and separation performance (Na2SO4 rejection of 94-96%) under hydraulic pressure-driven processes and eliminating internal concentration polarization in osmotic pressure-driven processes. Enhanced separation performances are achieved with water/Na2SO4 permselectivity of 13.5 L g-1 and high osmotic water permeability up to 1.41 L m-2 h-1 bar-1, providing 3-fold higher water/Na2SO4 permselectivity and 56-fold-higher water flux than polymer membranes for forward osmosis.

Characterizing the Adherence Profiles of Virulent Vibrio parahaemolyticus Isolates.

The human pathogen Vibrio parahaemolyticus is a leading cause of seafood-borne illness in the USA, and infections with V. parahaemolyticus typically result from eating raw or undercooked oysters. V. parahaemolyticus has been shown to be highly resistant to oyster depuration, suggesting that the bacterium possesses specific mechanisms or factors for colonizing oysters and persisting during depuration. In this study, we characterized eight different V. parahaemolyticus strains for differences in resistance to oyster depuration, biofilm formation, and motility. While each strain exhibited distinct phenotypes in the various assays, we determined that biofilm formation on abiotic surfaces, such as glass or plastic, does not directly correlate with bacterial retention in oysters during depuration. However, we did observe that the motility phenotype of a strain appeared to be a better indicator for persistence in the oyster. Further studies examining the molecular mechanisms underlying the observed colonization differences by these and other V. parahaemolyticus strains may provide beneficial insights into what critical factors are required for proficient colonization of the Pacific oyster.

Structural characterization and osteogenic bioactivity of a sulfated polysaccharide from pacific abalone (Haliotis discus hannai Ino).

Bone morphogenic protein-2 (BMP-2) is known to promote osteogenesis. To find novel adjuvants to enhance the activity of BMP-2, the present study investigated the structure BMP-2-induced osteogenic activity of a water-soluble polysaccharide from the gonad of pacific abalone (Haliotis discus hannai Ino) named AGSP. Through analysis of aldobiouronic acids released from AGSP, monosaccharide composition comparison of AGSP and its reduced product, and methylation analysis and NMR analysis of AGSP and its desulfated derivative, the main structure residue of AGSP was determined as →3)-GlcA(1→3)-Gal(1→ with sulfated branches comprised of prevelant Gal and minor Glc, and →4)-ß-GlcA(1→2)-α-Man(1→ residue was also found. AGSP possessed a sulfate content of 12.4% with a relative molecular weight of 6.6kDa. AGSP strengthened alkaline phosphatase activity induced by BMP-2 in a dose dependent manner at 10-200µg/mL with 425% enhancement being observed at 200µg/mL, indicating AGSP could be an adjuvant candidate to enhance osteogenic activity of BMP-2.

Degradation of Histamine by Lactobacillus plantarum Isolated from Miso Products.

Histamine is a toxic chemical and is the causative agent of food poisoning. This foodborne toxin may be degraded by the oxidative deamination activity of certain microorganisms. In this study, we isolated four histamine-degrading Lactobacillus plantarum bacteria from miso products. Among them, L. plantarum D-103 exhibited 100% degradation of histamine in de Man Rogosa Sharpe (MRS) broth containing 50 ppm of histamine after 24 h of incubation at 30°C. The optimal growth, histamine oxidase, and histamine-degrading activity of L. plantarum D-103 were observed in histamine MRS broth at pH 7.0, 3% NaCl, and 30°C. It also exhibited tolerance to broad ranges of pH (4 to 10) and salt concentrations (0 to 12%) in histamine MRS broth. Therefore, the histamine-degrading L. plantarum D-103 might be used as an additive culture to prevent histamine accumulation in miso products during fermentation.

Characterization of clinical Vibrio parahaemolyticus strains in Zhoushan, China, from 2013 to 2014.

Vibrio parahaemolyticus is recognized as major cause of foodborne illness of global public health concern. This study collected 107 strains of V. parahaemolyticus during active surveillance of diarrheal diseases in hospitals in Zhoushan during 2013 to 2014 and investigated their serotypes, virulence genes (tdh, trh, and orf8), antimicrobial resistance, and genotypes. The dominant serotypes of the 107 clinical strains were O3:K6, O4:K8, and O4:KUT with 87.9% and 3.7% of the strains carrying the virulence genes tdh and trh, respectively. Molecular typing by pulsed-field gel electrophoresis indicated divergence among the clinical strains. Most isolates were sensitive to the common antimicrobial agents used against the Vibrio species except ampicillin. We conclude that continuous surveillance of V. parahaemolyticus in diarrhea patients is a public health priority and is useful for conducting risk assessment of foodborne illnesses caused by V. parahaemolyticus.

Effect of spatial constraints on Hardy-Weinberg equilibrium.

Panmixia is a key issue in maintaining genetic diversity, which facilitates evolutionary potential during environmental changes. Additionally, conservation biologists suggest the importance of avoiding small or subdivided populations, which are prone to losing genetic diversity. In this paper, computer simulations were performed to the genetic drift of neutral alleles in random mating populations with or without spatial constraints by randomly choosing a mate among the closest neighbours. The results demonstrated that the number of generations required for the neutral allele to become homozygous (Th) varied proportionally to the population size and also strongly correlated with spatial constraints. The average Th for populations of the same size with spatial constraints was approximately one-and-a-half times longer than without constraints. With spatial constraints, homozygous population clusters formed, which reduced local diversity but preserved global diversity. Therefore, panmixia might be harmful in preserving the genetic diversity of an entire population. The results also suggested that the gene flow or gene exchange among the subdivided populations must be carefully processed to restrict diseases transmission or death during transportation and to monitor the genetic diversity. The application of this concept to similar systems, such as information transfer among peers, is also discussed.

Behavior of Salmonella and Listeria monocytogenes in Raw Yellowfin Tuna during Cold Storage.

Behavior of Salmonella and Listeria monocytogenes in raw yellowfin tuna during refrigeration and frozen storage were studied. Growth of Salmonella was inhibited in tuna during refrigerated storage, while L. monocytogenes was able to multiply significantly during refrigerated storage. Populations of Salmonella in tuna were reduced by 1 to 2 log after 12 days of storage at 5-7 °C, regardless levels of contamination. However, populations of L. monocytogenes Scott A, M0507, and SFL0404 in inoculated tuna (104-105 CFU/g) increased by 3.31, 3.56, and 3.98 log CFU/g, respectively, after 12 days of storage at 5-7 °C. Similar increases of L. monocytogenes cells were observed in tuna meat with a lower inoculation level (10²-10³ CFU/g). Populations of Salmonella and L. monocytogenes declined gradually in tuna samples over 84 days (12 weeks) of frozen storage at -18 °C with Salmonella Newport 6962 being decreased to undetectable level (<10 CFU/g) from an initial level of 10³ log CFU/g after 42 days of frozen storage. These results demonstrate that tuna meat intended for raw consumption must be handled properly from farm to table to reduce the risks of foodborne illness caused by Salmonella and L. monocytogenes.

Analysis of Apoptosis in Ultraviolet-Induced Sea Cucumber (Stichopus japonicus) Melting Using Terminal Deoxynucleotidyl-Transferase-Mediated dUTP Nick End-Labeling Assay and Cleaved Caspase-3 Immunohistochemistry.

The sea cucumber body wall melting phenomenon occurs under certain circumstances, and the mechanism of this phenomenon remains unclear. This study investigated the apoptosis in the ultraviolet (UV)-induced sea cucumber melting phenomenon. Fresh sea cucumbers (Stichopus japonicus) were exposed to UV radiation for half an hour at an intensity of 0.056 mW/cm(2) and then held at room temperature for melting development. The samples were histologically processed into formalin-fixed paraffin-embedded tissues. The apoptosis of samples was analyzed with the terminal deoxynucleotidyl-transferase-mediated dUTP nick end-labeling (TUNEL) assay and cleaved caspase-3 immunohistochemistry. The emergence of TUNEL-positive cells speeds up between 0.5 and 2 h after UV irradiation. Cleaved caspase-3 positive cells were obviously detected in sample tissues immediately after the UV irradiation. These results demonstrated that sea cucumber melting induced by UV irradiation was triggered by the activation of caspase-3 followed by DNA fragmentation in sea cucumber tissue, which was attributed to apoptosis but was not a consequence of autolysis activity.

Analysis of Vibrio vulnificus Infection Risk When Consuming Depurated Raw Oysters.

A beta Poisson dose-response model for Vibrio vulnificus food poisoning cases leading to septicemia was used to evaluate the effect of depuration at 15 °C on the estimated health risk associated with raw oyster consumption. Statistical variability sources included V. vulnificus level at harvest, time and temperature during harvest and transportation to processing plants, decimal reductions (SV) observed during experimental circulation depuration treatments, refrigerated storage time before consumption, oyster size, and number of oysters per consumption event. Although reaching nondetectable V. vulnificus levels (<30 most probable number per gram) throughout the year and a 3.52 SV were estimated not possible at the 95% confidence level, depuration for 1, 2, 3, and 4 days would reduce the warm season (June through September) risk from 2,669 cases to 558, 93, 38, and 47 cases per 100 million consumption events, respectively. At the 95% confidence level, 47 and 16 h of depuration would reduce the warm and transition season (April through May and October through November) risk, respectively, to 100 cases per 100 million consumption events, which is assumed to be an acceptable risk; 1 case per 100 million events would be the risk when consuming untreated raw oysters in the cold season (December through March).

Efficacy of low-temperature high hydrostatic pressure processing in inactivating Vibrio parahaemolyticus in culture suspension and oyster homogenate.

Culture suspensions of five clinical and five environmental Vibrio parahaemolyticus strains in 2% NaCl solution were subjected to high pressure processing (HPP) under various conditions (200-300MPa for 5 and 10 min at 1.5-20°C) to study differences in pressure resistance among the strains. The most pressure-resistant and pressure-sensitive strains were selected to investigate the effects of low temperatures (15, 5 and 1.5°C) on HPP (200 or 250MPa for 5 min) to inactivate V. parahaemolyticus in sterile oyster homogenates. Inactivation of V. parahaemolyticus cells in culture suspensions and oyster homogenates was greatly enhanced by lowering the processing temperature from 15 to 5 or 1.5°C. A treatment of oyster homogenates at 250MPa for 5 min at 5°C decreased the populations of V. parahaemolyticus by 6.2logCFU/g for strains 10290 and 100311Y11 and by >7.4logCFU/g for strain 10292. Decreasing the processing temperature of the same treatment to 1.5°C reduced all the V. parahaemolyticus strains inoculated to oyster homogenates to non-detectable (<10CFU/g) levels. Factors including pressure level, processing temperature and time all need to be considered for developing effective HPP for eliminating pathogens from foods. Further studies are needed to validate the efficacy of the HPP (250MPa for 5 min at 1.5°C) in inactivating V. parahaemolyticus cells in whole oysters.

Effects of frozen storage on survival of Staphylococcus aureus and enterotoxin production in precooked tuna meat.

This study investigated the survival of Staphylococcus aureus in precooked tuna meat for producing canned products during frozen storage (-20 ± 2 °C) as well as its growth and enterotoxin production at 35 to 37 °C after the storage. Samples (50 ± 5 g) of precooked albacore (loin, chunk, and flake) and skipjack (chunk and flake) tuna were inoculated with 5 enterotoxin-producing strains of S. aureus at a level of approximately 3.5 log CFU/g and individually packed in a vacuum bag after 3 h incubation at 35 to 37 °C. Vacuum-packed samples were stored in a freezer (-20 ± 2 °C) for 4 wk. The frozen samples were then thawed in 37 °C circulating water for 2 h and incubated at 35 to 37 °C for 22 h. Populations of S. aureus in all precooked tuna samples decreased slightly (<0.7 log CFU/g) after 4 wk of storage at -20 ± 2 °C, but increased rapidly once the samples were thawed and held at 35 to 37 °C. Total S. aureus counts in albacore and skipjack samples increased by greater than 3 log CFU/g after 6 and 8 h of exposure to 35 to 37 °C, respectively. All samples became spoiled after 10 h of exposure to 35 to 37 °C, while no enterotoxin was detected in any samples. However, enterotoxins were detected in albacore loin and other samples after 12 and 24 h of incubation at 35 to 37 °C, respectively. Frozen precooked tuna meat should be used for producing canned tuna within 6 to 8 h of thawing to avoid product spoilage and potential enterotoxin production by S. aureus in contaminated precooked tuna meat.

Persistence of Vibrio parahaemolyticus in the Pacific oyster, Crassostrea gigas, is a multifactorial process involving pili and flagella but not type III secretion systems or phase variation.

Vibrio parahaemolyticus can resist oyster depuration, suggesting that it possesses specific factors for persistence. We show that type I pili, type IV pili, and both flagellar systems contribute to V. parahaemolyticus persistence in Pacific oysters whereas type III secretion systems and phase variation do not.