Capsulated Strains Research Articles

Combined effect of artificial cervical disc replacement and facet tropism on the index-level facet joints: a finite element study

BackgroundArtificial Cervical Disc Replacement (ACDR) is an effective treatment for cervical degenerative disc diseases. However, clinical information regarding the facet joint alterations after ACDR was limited. Facet tropism is common in the sub-axial cervical spine. Our previous research indicated that facet tropism could lead to increased pressure on the cervical facet joints. This study aimed to assess the impact of facet tropism on the facet contact force and facet capsule stress after ACDR.MethodsA C2–T1 cervical finite element model was constructed from computed tomography (CT) scans of a 28-year-old male volunteer. Symmetrical, moderate asymmetrical (7 degrees tropism), and severe asymmetrical (14 degrees tropism) models were created at the C5/C6 level by altering the facet orientation at the C5-C6 level. The C5/C6 ACDR was simulated in the intact, moderate asymmetrical and severe asymmetrical models. A 75-N follower load with 1.0-Nm moments was applied to the top of C2 vertebra in the models to simulate flexion, extension, lateral bending, and axial rotation with the T1 vertebra fixed. The range of motions (ROMs) under all moments, facet contact forces (FCFs) and facet capsule strains were tested.ResultsIn the asymmetrical model, the right FCFs considerably increased under flexion, extension, right bending, left rotation, especially under right bending the right sided FCF of the severe asymmetrical model was about 5.44 times of the neutral position, and 3.14 times of the symmetrical model. and concentrated on the cephalad part of the facets. The facet capsule stresses on both sides remarkably increased under extension, lateral bending and right rotation. In the moderate and severe asymmetrical models, the capsule strain was greater on both sides of each position than in the symmetric model.ConclusionsThe face tropism increased facet contact force and facet capsule strain after ACDR, especially under extension, lateral bending, and rotation, and also could result in abnormal stress distribution on the facet joint surface and facet joint capsule. The results suggest that face tropism might be a risk factor for post-operative facet joint degeneration progression after ACDR. Facet tropism may be noteworthy when ACDR is considered as a surgical option.

Development of a finite element full spine model with active muscles for quantitatively analyzing sarcopenia effects on lumbar load

Background and objectiveThe musculoskeletal imbalance caused by disease is one of the most critical factors leading to spinal injuries, like sarcopenia. However, the effects of musculoskeletal imbalances on the spine are difficult to quantitatively investigate. Thus, a complete finite element spinal model was established to analyze the effects of musculoskeletal imbalance, especially concerning sarcopenia. MethodsA finite element spinal model with active muscles surrounding the vertebrae was established and validated from anatomic verification to the whole spine model in dynamic loading at multiple levels. It was then coupled with the previously developed neuromuscular model to quantitatively analyze the effects of erector spinae (ES) and multifidus (MF) sarcopenia on spinal tissues. The severity of the sarcopenia was classified into three levels by changing the physiological cross-sectional area (PCSA) of ES and MF, which were mild (60% PCSA of ES and MF), moderate (48% PCSA of ES and MF), and severe (36% PCSA of ES and MF). ResultsThe stress and strain levels of most lumbar tissues in the sarcopenia models were more significant than those of the normal model during spinal extension movement. The sarcopenia caused load concentration in several specific regions. The stress level of the L4-L5 intervertebral disc and L1 vertebra significantly increased with the severity of sarcopenia and showed relatively larger values than other segments. From the normal model to a severe sarcopenia model, the stress value of the L4-L5 intervertebral disc and L1 vertebra increased by 128% and 113%, respectively. The strain level of L5-S1 also inclined significantly with the severity of sarcopenia, and the relatively larger capsule strain values occurred at lower back segments from L3 to S1. ConclusionsIn summary, the validated spinal coupling model can be used for spinal injury risk analysis caused by musculoskeletal imbalance. The results suggested that sarcopenia can primarily lead to high injury risk of the L4-L5 intervertebral disc, L1 vertebrae, and L3-S1 joint capsule regarding significant stress or strain variance.

Different regulatory mechanisms of the capsule in hypervirulent Klebsiella pneumonia: "direct" wcaJ variation vs. "indirect" rmpA regulation.

Hypervirulent Klebsiella pneumoniae produce an increased amount of capsular substance and are associated with a hypermucoviscous phenotype. Capsule production is regulated by capsular regulatory genes and capsular gene cluster variations. In the present study, we focus on the effect of rmpA and wcaJon capsule biosynthesis. Phylogenetic trees were constructed to analyze wcaJ and rmpA sequence diversity in different serotypes hypervirulent strains. Then mutant strains (K2044ΔwcaJ, K2044K1wcaJ, K2044K2wcaJand K2044K64wcaJ) were used to verify the effects of wcaJ and its diversity on capsule synthesis and strain virulence. Furthmore, the role of rmpA in capsular synthesis and its mechanisms were detected in K2044ΔrmpA strain. RmpA sequences are conversed in different serotypes. And rmpA promoted the production of hypercapsules by simultaneously acting on three promoters in cps cluster. Whereas wcaJ, its sequences are different in different serotypes, and its loss result in the termination of capsular synthesis. Moreover, the results verified that K2 wcaJ could form hypercapsule in K2044 strains (K1 serotype), but K64 wcaJ could not. The interaction of multiple factors is involved in capsule synthesis, including wcaJ and rmpA. RmpA, an known conserved capsular regulator gene, acts on cps cluster promoters to promote the production of the hypercapsule. WcaJ as initiating enzyme of CPS biosynthesis, its presence determines the synthesis of capsule. Besides, different from rmpA, wcaJ sequence consistency is limited to the same serotype, which cause wcaJ functioning in different serotype strains with sequence recognition specificity.

Assessment of Capsule Related Antibiotic Resistance of Pseudomonas aeruginosa Isolated from Patients Attending some Hospitals in Kano State, Nigeria

Resistance to antimicrobial substances by pathogenic microorganisms is on the rise since the beginning of modern medicine. Mechanisms; enzymatic degradation, loss of target and antimicrobial interception are employed by bacteria in resisting antimicrobials. Bacterial capsule is a gelatinous outer layer surrounding a bacterial cell composed primarily of polysaccharide. This study assessed the role that the capsule of Pseudomonas aeruginosa played in carbapenem resistance. P. aeruginosa was isolated from 49 (16%) samples out of 300 samples screened. India ink staining technique was used to detect the capsule. Capsule was detected in 42 (86%) of the isolates whereas 7 (14%) of the isolates were non-capsulated. All the capsulated strains were resistant to the antibiotics (Meropenem, Doripenem and Imipenem) whereas the non-capsulated strains were susceptible to the antibiotics. The results of this study however suggest the involvement of capsule in the resistance of the antibiotics employed.

Typeable ampicillin-resistant Haemophilus influenzae strains in Tunisian's children.

Background:Here, we report the frequency of capsulated ampicillin-resistant Haemophilus influenzae strains isolated from children in Tunisia, particularly capsular serotype b, by polymerase chain reaction (PCR) to determine the molecular mechanisms underlying ampicillin resistance.Methods:We considered 22 capsulated H influenzae strains selected from a series of 91 ampicillin-resistant H influenzae strains isolated from children between 2010 and 2011 in Tunisia. The capsular serotypes of these strains were identified by slide agglutination and PCR.Results:By PCR, 19 (20.88%) serotype b, 1 (1.1%) serotype a, 2 (2.2%) serotypes d and f and 69 (75.82%) non-typeable strains were found among the 91 ampicillin-resistant H influenzae strains. 100% of the assumption between the consequences of antigenic examinations and PCR was found. The serotype b strains showed biotypes I, II, III, IV, VI, and VIII. The other capsulated strains showed biotypes IV and VIII. Thirteen of the serotype b strains created β-lactamase (14.28%). The 19 serotype b ampicillin-resistant H influenzae strains were subdivided into 3 bunches as indicated: The gathering of the β-lactamase positive, ampicillin-resistant where 11 strains (57.89%) were β-lactamase positive blaTEM-1 (+) and ftsI (+). The second gathering of the β-lactamase negative, ampicillin-resistant strains, where 6 isolates (31.58%) were β-lactamase negative blaTEM-1 (−) and ftsI (−), and lastly, the gathering of the β-lactamase positive, amoxicillin-clavulanate resistant where 2 isolates (10.52%) were β-lactamase positive blaTEM-1 (+) and ftsI (−).Conclusion:PCR should be used in our country because it may contribute to decreasing the probability of transmission of these strains, especially those showing the two mechanisms of resistance among children in Tunisia.

Comparative pangenome analysis of capsulated Haemophilus influenzae serotype f highlights their high genomic stability

Haemophilus influenzae is an opportunistic pathogen adapted to the human respiratory tract. Non-typeable H. influenzae are highly heterogeneous, but few studies have analysed the genomic variability of capsulated strains. This study aims to examine the genetic diversity of 37 serotype f isolates from the Netherlands, Portugal, and Spain, and to compare all capsulated genomes available on public databases. Serotype f isolates belonged to CC124 and shared few single nucleotide polymorphisms (SNPs) (n = 10,999), but a high core genome (> 80%). Three main clades were identified by the presence of 75, 60 and 41 exclusive genes for each clade, respectively. Multi-locus sequence type analysis of all capsulated genomes revealed a reduced number of clonal complexes associated with each serotype. Pangenome analysis showed a large pool of genes (n = 6360), many of which were accessory genome (n = 5323). Phylogenetic analysis revealed that serotypes a, b, and f had greater diversity. The total number of SNPs in serotype f was significantly lower than in serotypes a, b, and e (p < 0.0001), indicating low variability within the serotype f clonal complexes. Capsulated H. influenzae are genetically homogeneous, with few lineages in each serotype. Serotype f has high genetic stability regardless of time and country of isolation.

Exploring the influence of impact severity and posture on vertebral joint mechanics in an in-vitro porcine model

To date, no in vitro studies have been conducted to explore lumbar soft tissue injury potential and altered mechanical properties from exposure to impact forces. After a motor vehicle collision (MVC), the cause of reported acute onset low back pain is difficult to associate with potential soft tissue strain injury sites including the facet joint and innervated facet joint capsule ligament (FJC). Thus, the purpose of this investigation was to quantify intervertebral anterior-posterior (AP) translation and facet joint capsule strain under varying postures and impact severities. Seventy-two porcine spinal units were exposed to three levels of impact severity (4 g, 8 g, 11 g), and posture (Neutral, Flexion, Extension). Impacts were applied using a custom-built impact track that replicated parameters experienced in low to moderate speed rear-end MVCs. Flexion-extension and anterior-posterior shear neutral zone testing were completed prior to impact. AP intervertebral translation and the strain tensor of the facet capsule ligament were measured during impacts. A significant main effect of collision severity was observed for peak AP intervertebral translation (4 g—2.8 ±0.53 mm; 8 g—6.4 ±2.9 mm; 11 g—8.3 ±0.45 mm) and peak FJC shear strain (2.37% strain change from 4 g to 11 g impact severity). Despite observed main effects of impact severity, no influence of posture was observed. This lack of influence of posture and small FJC strain magnitudes suggest that the FJC does not appear to undergo injurious or permanent mechanical changes in response to low-to-moderate MVC impact scenarios.

Rapid identification of capsulated Acinetobacter baumannii using a density-dependent gradient test

BackgroundGram-negative bacterial capsules are associated with production of carbohydrates, frequently resulting in a mucoid phenotype. Infections caused by capsulated or mucoid A. baumannii are associated with increased clinical severity. Therefore, it is clinically and epidemiologically important to identify capsulated A. baumannii. Here, we describe a density-dependent gradient test to distinguish between capsulated and thin/non-capsulated A. baumannii.ResultsThirty-one of 57 A. baumannii isolates displayed a mucoid phenotype. The density-dependent gradient test was comprised of two phases, with silica concentrations of 30% (top phase) and 50% (bottom phase). Twenty-three isolates migrated to the bottom phase, indicating thin or non-capsulated strains, and 34 migrated to the top phase, suggesting strains suspected to be capsulated. There was agreement between the mucoid and the non-mucoid phenotypes and the density-dependent gradient test for all but three isolates. Total carbohydrates extracted from strains suspected to be capsulated were significantly higher. Transmission electron microscopy confirmed the presence of a capsule in the six representative strains suspected to be capsulated.ConclusionsThe density-dependent gradient test can be used to verify capsule presence in mucoid-appearing A. baumannii strains. Identifying capsulated strains can be useful for directing infection control measures to reduce the spread of hypervirulent strains.

Serotyping strains of Haemophilus influenzae isolated from clinical specimens and investigating their susceptibilities towards certain antimicrobials

Aim: Haemophilus influenzae is an agent of severe pediatric infections, and a significant cause of mortality and morbidity in children. Its capsulated strains are grouped from ‘a’ to ‘f’ in six groups, the most invasive among which is Type B. The other serotypes are also capable of causing severe infections and invasive diseases. Antibiotic sensitivity testing proves to be useful in directing course of treatment in view of the beta-lactamase-negative and ampicillin-resistant strains as well as the ever-increasing production of beta-lactamase. In this study it has been aimed to serotype Haemophilus influenzae strains isolated from various clinical specimens, and to investigate their status of beta-lactamase production and susceptibilities towards antimicrobials. Materials and Methods: 100 samples of H. influenzae proliferating in various clinical specimens were included in the study. The bacteria were identified according to their requirements of X and V factors, or through the usage of API NH. Detection of beta-lactamase production was made with nitrocefin disks. Serotyping was performed via slide agglutination method with specific antiserums. Susceptibilities towards various antimicrobials were specified with gradient test in Haemophilus Test Medium. Results: 59 of the specimens were sputum, 23 were nasotracheal aspirates, 3 were bronchoalveolar lavages, 7 were middle ear fluids, 3 were cerebrospinal fluids, 2 were eye fluids, 2 were pleural fluids, and 1 was a blood sample. The ranking was made without keeping age groups in regard and turned out as a>b>d>THi>c>e. Serotype f was not found among the strains. Type b was isolated from all of sterile body cavity fluids. Beta-lactamase positivity was found as 2% and the BLNAR ratio was found as 2%. All were found as susceptible towards cefotaxime and siprofloxacin while 66.6% were susceptible towards azithromycin, and 93% were susceptible towards TMP-SMX. Conclusion: It will be appropriate to perform standardized susceptibility tests in routine in order to monitor antimicrobial resistance. Serotyping is important as inclusion of H. influenzae type b in vaccination programs may cause an increase in infections with non-Hib isolates.

Strain of the facet joint capsule during rotation and translation range-of-motion tests: an in vitro porcine model as a human surrogate

BACKGROUND CONTEXTPrior data about the modulating effects of lumbar spine posture on facet capsule strains are limited to small joint deviations. Knowledge of facet capsule strain during rotational and translational intervertebral joint motion (ie, large joint deviations) under physiological loading could be useful as it may help explain why visually normal lumbar spinal joints become painful. PURPOSEThis study quantified the strain tensor of the facet capsule during rotation and translation range-of-motion tests. STUDY DESIGN/SETTINGStrain was calculated in isolated porcine functional spinal units. Following a preload, each specimen underwent a flexion/extension rotation (F/E) followed by an anterior/posterior translation (A/P) range-of-motion test while under a 300 N compression load. METHODSTwenty porcine spinal units (10 C3–C4, 10 C5–C6) were tested. Joint flexion/extension was imposed by applying a ±8 Nm moment at a rate of 0.5°/s, and translation was facilitated by loading the caudal vertebra with a ±400 N shear force at a rate of 0.2 mm/s. Points were drawn on the exposed capsule surface and their coordinates were optically tracked throughout each test. Strain was calculated as the displacement of the point configuration with respect to the configuration in a neutral joint position. RESULTSCompared to a neutral posture, superior-inferior strain increased and decreased systematically during flexion and extension, respectively. Posterior displacement of the caudal vertebra by more than 1.3 mm was associated with negative strains, which was significantly lower than the +4.6% strain observed during anterior displacement (p≥.199). The shear strain associated with anterior translation was, on average, −1.1% compared to a neutral joint posture. CONCLUSIONSThese results demonstrate that there is a combination of strain types within the facet capsule when spinal units are rotated and translated. The strains documented in this study did not reach the thresholds associated with nociception. CLINICAL RELEVANCEThe magnitude of flexion-extension rotation and anterior-translation may glean insight into the facet capsule deformation response under low compression (300 N) loading scenarios. Further, intervertebral joint motion alone, even under low compression loading, does not appear to initiate a clinically relevant pain response in the lumbar facet capsule of a nondegenerated spinal joint.

A biomechanical analysis of prognathous and orthognathous insect head capsules: evidence for a many-to-one mapping of form to function.

Insect head shapes are remarkably variable, but the influences of these changes on biomechanical performance are unclear. Among 'basal' winged insects, such as dragonflies, mayflies, earwigs and stoneflies, some of the most prominent anatomical changes are the general mouthpart orientation, eye size and the connection of the endoskeleton to the head. Here, we assess these variations as well as differing ridge and sclerite configurations using modern engineering methods including multibody dynamics modelling and finite element analysis in order to quantify and compare the influence of anatomical changes on strain in particular head regions and the whole head. We show that a range of peculiar structures such as the genal/subgenal, epistomal and circumocular areas are consistently highly loaded in all species, despite drastically differing morphologies in species with forward-projecting (prognathous) and downward-projecting (orthognathous) mouthparts. Sensitivity analyses show that the presence of eyes has a negligible influence on head capsule strain if a circumocular ridge is present. In contrast, the connection of the dorsal endoskeletal arms to the head capsule especially affects overall head loading in species with downward-projecting mouthparts. Analysis of the relative strains between species for each head region reveals that concerted changes in head substructures such as the subgenal area, the endoskeleton and the epistomal area lead to a consistent relative loading for the whole head capsule and vulnerable structures such as the eyes. It appears that biting-chewing loads are managed by a system of strengthening ridges on the head capsule irrespective of the general mouthpart and head orientation. Concerted changes in ridge and endoskeleton configuration might allow for more radical anatomical changes such as the general mouthpart orientation, which could be an explanation for the variability of this trait among insects. In an evolutionary context, many-to-one mapping of strain patterns onto a relatively similar overall head loading indeed could have fostered the dynamic diversification processes seen in insects.

Anterior lens capsule strains during simulated accommodation in porcine eyes

Experimental protocols have been developed to measure the spatial variation of the mechanical strains induced in the lens capsule during ex vivo lens stretching. The paper describes the application of these protocols to porcine lenses. The deformations and mechanical strains developed in the anterior capsule during each experiment were determined using full field digital image correlation techniques, by means of a speckle pattern applied to the lens surface. Several speckling techniques and illumination methods were assessed before a suitable combination was found. Additional data on the cross section shape of the anterior lens surface were obtained by Scheimpflug photography, to provide a means of correcting for lens curvature effects in the determination of the strains developed in the plane of the capsule. The capsule strains in porcine lenses exhibit non-linear behaviour, and hysteresis during loading and unloading. Peripheral regions experience higher magnitude strains than regions near the lens pole. The paper demonstrates the successful application of a procedure to make direct measurements of capsule strains simultaneously with ex vivo radial lens stretching. This experimental technique is applicable to future investigations on the mechanical characteristics of human lenses.

Collagen organization regulates stretch-initiated pain-related neuronal signals in vitro: Implications for structure-function relationships in innervated ligaments.

Injury to the spinal facet capsule, an innervated ligament with heterogeneous collagen organization, produces pain. Although mechanical facet joint trauma activates embedded afferents, it is unclear if, and how, the varied extracellular microstructure of its ligament affects sensory transduction for pain from mechanical inputs. To investigate the effects of macroscopic deformations on afferents in collagen matrices with different organizations, an in vitro neuron-collagen construct (NCC) model was used. NCCs with either randomly organized or parallel aligned collagen fibers were used to mimic the varied microstructure in the facet capsular ligament. Embryonic rat dorsal root ganglia (DRG) were encapsulated in the NCCs; axonal outgrowth was uniform and in all directions in random NCCs, but parallel in aligned NCCs. NCCs underwent uniaxial stretch (0.25 ± 0.06 strain) corresponding to sub-failure facet capsule strains that induce pain. Macroscopic NCC mechanics were measured and axonal expression of phosphorylated extracellular signal-regulated kinase (pERK) and the neurotransmitter substance P (SP) was assayed at 1 day to assess neuronal activation and nociception. Stretch significantly upregulated pERK expression in both random and aligned gels (p < 0.001), with the increase in pERK being significantly higher (p = 0.013) in aligned than in random NCCs. That increase likely relates to the higher peak force (p = 0.025) and stronger axon alignment (p < 0.001) with stretch direction in the aligned NCCs. In contrast, SP expression was greater in stretched NCCs (p < 0.001) regardless of collagen organization. These findings suggest that collagen organization differentially modulates pain-related neuronal signaling and support structural heterogeneity of ligament tissue as mediating sensory function. © 2017 Orthopaedic Research Society. Published by Wiley Periodicals, Inc. J Orthop Res 36:770-777, 2018.

Porphyromonas gingivalis Capsule-Mediated Coaggregation as a Virulence Factor in Mixed Infection With Fusobacterium nucleatum.

Porphyromonas gingivalis (Pg) capsule enables evasion from phagocytosis, invasion of keratinocytes, and bacterial survival. In mixed infection, the capsule also participates in coaggregation, which may lead to characteristic virulence not present in the monoinfection. The aim of this study is to evaluate the role of Pg capsule as a virulence factor in coaggregated mixed infection with Fusobacterium nucleatum (Fn). Mixed infections containing Fn and non-capsulated or capsulated strains of Pg were compared with the same infection with lactose as coaggregation inhibitor. Murine experimental periodontitis was used to assess disease severity. Primary polymorphonuclear leukocytes and keratinocytes were used to examine phagocytosis and bacterial invasion, respectively. Mixed infection with capsulated Pg augmented alveolar bone loss compared with that of mixed infection with non-capsulated Pg. Addition of lactose led to attenuation of bone loss in the capsulated mixed infection and to intensification of bone loss in the non-capsulated mixed infection. In the latter mixed infection, Fn evaded phagocytosis, whereas in the capsulated mixed infection Pg displayed a greater capacity for invasion of keratinocytes. Pg capsule was found to serve as a unique virulence factor in mixed infection with Fn. Capsule-dependent coaggregation led to augmented invasion of Pg and may be responsible for the severity of disease after mixed infection with Fn.

Upper Cervical Spine Loading Simulating a Dynamic Low-Speed Collision Significantly Increases the Risk of Pain Compared to Quasi-Static Loading With Equivalent Neck Kinematics.

Dynamic cervical spine loading can produce facet capsule injury. Despite a large proportion of neck pain being attributable to the C2/C3 facet capsule, potential mechanisms are not understood. This study replicated low-speed frontal and rear-end traffic collisions in occiput-C3 human cadaveric cervical spine specimens and used kinematic and full-field strain analyses to assess injury. Specimens were loaded quasi-statically in flexion and extension before and after dynamic rotation of C3 at 100 deg/s. Global kinematics in the sagittal plane were tracked at 1 kHz, and C2/C3 facet capsule full-field strains were measured. Dynamic loading did not alter the kinematics from those during quasi-static (QS) loading, but maximum principal strain (MPS) and shear strain (SS) were significantly higher (p = 0.028) in dynamic flexion than for the same quasi-static conditions. The full-field strain analysis demonstrated that capsule strain was inhomogeneous, and that the peak MPS generally occurred in the anterior aspect and along the line of the C2/C3 facet joint. The strain magnitude in dynamic flexion continued to rise after the rotation of C3 had stopped, with a peak MPS of 12.52 ± 4.59% and a maximum SS of 5.34 ± 1.60%. The peak MPS in loading representative of rear-end collisions approached magnitudes previously shown to induce pain in vivo, whereas strain analysis using linear approaches across the facet joint was lower and may underestimate injury risk compared to full-field analysis. The time at which peak MPS occurred suggests that the deceleration following a collision is critical in relation to the production of injurious strains within the facet capsule.

Molecular Basis for the Recognition of Higher Fullerenes into Ureidopyrimidinone-Cyclotriveratrylene Self-Assembled Capsules.

Fullerenes C60 , C70 , and C84 may be readily encaged within a hydrogen-bonded dimeric capsule, based on two concave cyclotriveratrylene (CTV) scaffolds, each containing three self-complementary 2-ureido-4-[1H]-pyrimidinone (UPy) subunits. NMR spectroscopy and circular dichroism studies, complemented by dispersion-corrected DFT calculations, are reported with the aim of characterizing such capsule-fullerene complexes both structurally and energetically. Six fullerenes are considered: in agreement with experiments, calculations find that encapsulation is most favorable for C84 (on a par with C90 ), and follows the trend C60 <C70 <C76 <C78 <C84 ≈C90 . As also found by NMR spectroscopy, the most stable capsules are formed by monomers of identical stereochemistry with UPy and the OMe groups of CTV in a mutual anti orientation and UPy-CTV ethylene linkers within the capsular surface plane. Up to C78 , encapsulation is favored by increasing host-guest contacts, but the trend is thereafter increasingly inhibited by growing capsule strain.

Comparative analysis of the immunologic response induced by the Sterne 34F2 live spore Bacillus anthracis vaccine in a ruminant model

The Sterne 34F2 live spore vaccine (SLSV) developed in 1937 is the most widely used veterinary vaccine against anthrax. However, literature on the immunogenicity of this vaccine in a target ruminant host is scarce. In this study, we evaluated the humoral response to the Bacillus anthracis protective antigen (rPA), a recombinant bacillus collagen-like protein of anthracis (rBclA), formaldehyde inactivated spores (FIS) prepared from strain 34F2 and a vegetative antigen formulation prepared from a capsule and toxin deficient strain (CDC 1014) in Boer goats. The toxin neutralizing ability of induced antibodies was evaluated using an in vitro toxin neutralization assay. The protection afforded by the vaccine was also assessed in vaccinates. Anti-rPA, anti-FIS and lethal toxin neutralizing titres were superior after booster vaccinations, compared to single vaccinations. Qualitative analysis of humoral responses to rPA, rBclA and FIS antigens revealed a preponderance of anti-FIS IgG titres following either single or double vaccinations with the SLSV. Antibodies against FIS and rPA both increased by 350 and 300-fold following revaccinations respectively. There was no response to rBclA following vaccinations with the SLSV. Toxin neutralizing titres increased by 80-fold after single vaccination and 700-fold following a double vaccination. Lethal challenge studies in naïve goats indicated a minimum infective dose of 36 B. anthracis spores. Single and double vaccination with the SLSV protected 4/5 and 3/3 of goats challenged with>800 spores respectively. An early booster vaccination following the first immunization is suggested in order to achieve a robust immunity. Results from this study indicate that this crucial second vaccination can be administered as early as 3 months after the initial vaccination.

Comparison of PCR-based methods for the simultaneous detection of Neisseria meningitidis, Haemophilus influenzae, and Streptococcus pneumoniae in clinical samples

BackgroundSeveral in-house PCR-based assays have been described for the detection of bacterial meningitis caused by Neisseria meningitidis, Streptococcus pneumoniae, and Haemophilus influenzae from clinical samples. PCR-based methods targeting different bacterial genes are frequently used by different laboratories worldwide, but no standard method has ever been established. The aim of our study was to compare different in-house and a commercial PCR-based tests for the detection of bacterial pathogens causing meningitis and invasive disease in humans. MethodsA total of 110 isolates and 134 clinical samples (99 cerebrospinal fluid and 35 blood samples) collected from suspected cases of invasive disease were analyzed. Specific sets of primers frequently used for PCR-diagnosis of the three pathogens were used and compared with the results achieved using the multiplex approach described here. Several different gene targets were used for each microorganism, namely ctrA, crgA and nspA for N. meningitidis, ply for S. pneumoniae, P6 and bexA for H. influenzae. ResultsAll used methods were fast, specific and sensitive, while some of the targets used for the in-house PCR assay detected lower concentrations of genomic DNA than the commercial method. An additional PCR reaction is described for the differentiation of capsulated and non-capsulated H. influenzae strains, the while commercial method only detects capsulated strains. ConclusionsThe in-house PCR methods here compared showed to be rapid, sensitive, highly specific, and cheaper than commercial methods. The in-house PCR methods could be easily adopted by public laboratories of developing countries for diagnostic purposes. The best results were achieved using primers targeting the genes nspA, ply, and P6 which were able to detect the lowest DNA concentrations for each specific target.

A pre-tensioned finite element model of ocular accommodation and presbyopia

Accommodation is the ability of the eye to dynamically alter its optical power. Presbyopia is the progressive loss of this ability with age. We have developed the first pre-tensioned finite element model of accommodation and used it to address the question of whether increasing lens stiffness alone is sufficient to cause presbyopia. Intrinsic resting forces and accommodative displacement were calibrated for a 20 year-old lens’ stiffness over a 7 D optical power change. Accommodation was simulated by displacing the distal zonule attachments towards the optical axis, mimicking the movement of the ciliary body when the ciliary muscle contracts. The optical power, capsule strain, and radial zonule forces were extracted for one full cycle of accommodation. These responses were compared for lenses having elastic moduli corresponding to 14, 20, 39.5, and 62.7-year-old human lenses. Accommodative power decreased from 20 to 62.5 years of age from 7 to 2 D. This change was less than in experiments where changes in lens power go to zero around age 50. Thus, lens elasticity contributes significantly to presbyopia but is not the sole cause. Lens diameter and thickness changes were within 20 % of reported values, indicating that the present model yields a good approximation of the changes in lens shape during accommodation.

Identification of a Monoclonal Antibody Against Pneumococcal Pilus 1 Ancillary Protein Impairing Bacterial Adhesion to Human Epithelial Cells.

The adhesion of Streptococcus pneumoniae is a key step during colonization of human respiratory tract mucosae. Here we demonstrate that pneumococcal type I pilus significantly increases the adhesiveness of poorly adhering highly capsulated strains in vitro. Interestingly, preincubation of bacteria with antibodies against the major pilus backbone subunit (RrgB) or the adhesin component (RrgA) impaired pneumococcal association to human epithelial cells. Screening for anti-RrgA monoclonal antibodies specifically affecting the adhesive capacity of S. pneumoniae led to the identification of the monoclonal 11B9/61 antibody, which greatly reduced pilus-dependent cell contact. Proteomic-based epitope mapping of 11B9/61 monoclonal antibody revealed a well-exposed epitope on the D2 domain of RrgA as the target of this functional antibody. The data presented here confirm the importance of pilus I for S. pneumoniae pathogenesis and the potential use of antipilus antibodies to prevent bacterial colonization.