Capsule Size Research Articles

A Quorum-Sensing System That Regulates Streptococcus pneumoniae Biofilm Formation and Surface Polysaccharide Production.

Despite vaccines, Streptococcus pneumoniae kills more than a million people yearly. Thus, understanding how pneumococci transition from commensals to pathogens is particularly relevant. Quorum sensing regulates collective behaviors and thus represents a potential driver of commensal-to-pathogen transitions. Rgg/small hydrophobic peptide (SHP) quorum-sensing systems are widespread in streptococci, yet they remain largely uncharacterized in S.pneumoniae. Using directional transcriptome sequencing, we show that the S.pneumoniae D39 Rgg0939/SHP system induces the transcription of a single gene cluster including shp and capsule gene homologs. Capsule size measurements determined by fluorescein isothiocyanate-dextran exclusion allowed assignment of the system to the regulation of surface polysaccharide expression. We found that the SHP pheromone induced exopolysaccharide expression in R36A, an unencapsulated derivative of D39. In the encapsulated parent strain, overexpression of the Rgg system resulted in a mutant with increased capsule size. In line with previous studies showing that capsule expression is inversely associated with biofilm formation, we found that biofilm formed on lung epithelial cells was decreased in the overexpression strain and increased in an rgg deletion mutant. Although no significant differences were observed between D39 and the rgg deletion mutant in a mouse model of lung infection, in competitive assays, overexpression reduced fitness. This is the first study to reveal a quorum-sensing system in streptococci that regulates exopolysaccharide synthesis from a site distinct from the original capsule locus. IMPORTANCE Quorum sensing regulates bacterial social behaviors by production, secretion, and sensing of pheromones. In this study, we characterized a new quorum-sensing system of the Rgg/SHP class in S.pneumoniae D39. The system was found to directly induce the expression of a single gene cluster comprising the gene for the SHP pheromone and genes with putative functions in capsule synthesis. Capsule size, as measured by dextran exclusion, was increased by SHP exposure in R36A, an unencapsulated derivative of D39. In the encapsulated parent strain, overexpression of the gene cluster increased capsule size, supporting the role of Rgg/SHP in the synthesis of surface polysaccharides. Further, we found that biofilm formation on epithelial cells was reduced by overexpression of the system and increased in a mutant with an rgg deletion. Placing surface polysaccharide expression under quorum-sensing regulation may enable S.pneumoniae to tune interactions with the host and other bacteria in accordance with environmental and cell density conditions.

Local Variation in Egg-Capsule Size in New England Populations of Nucella lapillus (Atlantic Dogwhelk)

Nucella lapillus (Atlantic Dogwhelk) deposits egg capsules on solid, intertidal substrates across the North Atlantic. This study investigated whether regional geographic variation or local wave-exposure affect the size of Dogwhelk egg capsules. Over 3 years, we evaluated whether Dogwhelks from wave-exposed and wave-protected sites in Massachusetts and mid-coast Maine differed in their egg-capsule size. Our results indicate that egg capsules collected from wave-exposed sites in Maine were smaller than egg capsules from wave-protected sites in Maine, but the size of egg capsules from Massachusetts did not vary with wave-exposure. These patterns in egg-capsule size coincide with Dogwhelk size from the same sites. Despite the positive correlation between the sizes of adult Dogwhelks and the egg capsules collected, wave-protected Dogwhelks from Massachusetts showed plasticity in the size of egg capsules produced but those from Maine did not. The Massachusetts Dogwhelk's greater plasticity in egg-capsule size highlights important local variation in control of reproductive investment and may accommodate fluctuations in desiccation stress and future climate change.

Structural Control of Carbon-Free Catalyst Layer Using Connected Platinum-Iron Nanoparticle Catalyst for Improved Mass-Transport in Polymer Electrolyte Fuel Cells

We have developed the connected platinum-iron (PtFe) nanoparticle catalyst with a porous hollow capsule structure as a carbon-free catalyst-layer material for polymer electrolyte fuel cells (PEFCs). [1,2] As shown in Fig. 1A, this catalyst consists of a nanosized beaded network by the fusion of PtFe nanoparticles with a crystallite size of 6~7 nm and an L10type chemically-ordered (face centered tetragonal) structure. The beaded metal network is electrically conductive, enabling the removal of carbon supports form a catalyst layer. The carbon-free cathode catalyst layer using a connected PtFe catalyst exhibits high durability against start-stop cycles, because of elimination of carbon corrosion problems. In addition, the specific activity of a connected PtFe catalyst for oxygen-reduction-reaction (ORR) is about 9 times enhanced, compared with that of a commercial Pt-nanoparticle catalyst supported on carbon black (Pt/C). On the other hand, inside a cathode catalyst layer, complex phenomena occur during a fuel-cell operation, such as ORR on catalyst surface, diffusion of oxygen molecules, proton conduction via ionomers, etc. Thus, a high performance of PEFC requires not only high ORR activity and stability but also lower resistance of oxygen mass transport. In this study, the following structural effects on fuel-cell performances for the carbon-free catalyst layers using connected PtFe catalysts were investigated; (i) inner-space in hollow capsule, and (ii) inter-space between capsules (Fig. 1B). The effect of inner-space in hollow capsule was discussed by the comparison of the connected PtFe catalysts with hollow structure or on solid (silica) support. The inter-space between capsules was controlled by the use of different capsule sizes (capsule size = 300 nm, or 500 nm in diameter). The electrochemical analyses of the prepared catalyst layers revealed that the hollow structure and the enlargement of inter-space between capsules improved the IV performance, especially at the high current density region, implying an improved oxygen mass-transport due to a sufficient space for oxygen diffusion in the catalyst layer and/or effective removal of liquid water from the catalyst layer. Moreover, in this study, the structural effect of gas-diffusion layer on fuel-cell performances was also investigated. It was found that the gas diffusion layer with higher gas permeability would facilitate the removal of liquid water from the capsule catalyst layer, resulting in a lower oxygen-transport resistance. In this presentation, by means of structural and electrochemical analyses of the capsule catalyst layers, the relationship between the catalyst-layer nanostructures and the fuel-cell performances will be discussed in details. The knowledge obtained in this study will provide new insights into the design of a carbon-free catalyst layer to achieve an enhanced PEFC performance. [1] T. Tamaki, H. Kuroki, S. Ogura, T. Fuchigami, Y. Kitamoto, and T. Yamaguchi, Energy Environ. Sci., 8, 3545-3549 (2015). [2] H. Kuroki, T. Tamaki, and T. Yamaguchi, J. Electrochem. Soc., 163(8), F927-F932 (2016). Figure 1

Biology and Rearing of Ectomyeolis ceratoniae Zeller (Lepidoptera: Pyralidae), Carob Moth, a Pest of Multiple Crops in South Africa

Ectomyeolis ceratoniae Zeller (Lepidoptera: Pyralidae), carob moth, is a pest of several crops in South Africa. A laboratory culture was established from field-collected larvae infesting mummified pecan nuts. Biological parameters of larvae reared on an artificial diet were measured. The insect goes through five larval instars, and the head capsule sizes of the five instars were determined to be ≤0.34 mm, 0.35–0.64 mm, 0.65–0.94 mm, 0.95–1.14 mm and ≥0.15 mm for the five instars, respectively. The insect was reared individually and communally in glass vials, the latter to develop a mass-rearing technique. Developmental time from neonate to pupa was significantly slower when larvae were individually reared (38.18 ± 1.2 days) compared to when they were communally reared (24.6 ± 0.65 days).Amicrosporidian infection (Nosema sp.) was recorded in the culture, causing significantly (F1,6 = 14.99, P = 0.0082) higher mortality of communally reared larvae (76.25 % ± 11.87) than individually reared larvae (24.9 % ± 9.6).

LI-RADS major features: CT, MRI with extracellular agents, and MRI with hepatobiliary agents.

The Liver Imaging Reporting and Data System (LI-RADS) was designed to standardize the interpretation and reporting of observations seen on studies performed in patients at risk for development of hepatocellular carcinoma (HCC). The LI-RADS algorithm guides radiologists through the process of categorizing observations on a spectrum from definitely benign to definitely HCC. Major features are the imaging features used to categorize observations as LI-RADS 3 (intermediate probability of malignancy), LIRADS 4 (probably HCC), and LI-RADS 5 (definite HCC). Major features include arterial phase hyperenhancement, washout appearance, enhancing capsule appearance, size, and threshold growth. Observations that have few major criteria are assigned lower categories than those that have several, with the goal of preserving high specificity for the LR-5 category of Definite HCC. The goal of this paper is to discuss LI-RADS major features, including definitions, rationale for selection as major features, and imaging examples.

Analytical Model for the Probability Characteristics of a Crack Penetrating Capsules in Capsule-Based Self-Healing Cementitious Materials

Autonomous crack healing using pre-embedded capsules containing healing agent is becoming a promising approach to restore the strength of damaged structures. In addition to the material properties, the size and volume fraction of capsules influence crack healing in the matrix. Understanding the crack and capsule interaction is critical in the development and design of structures made of capsule-based self-healing materials. Continuing our previous study, in this contribution a more practical rupturing mode of capsules characterizing the rupturing manner of capsules fractured by cracks in cementitious materials is presented, i.e., penetrating mode. With the underlying assumption that a crack penetrating capsules undoubtedly leads to crack healing, geometrical probability theory is employed to develop the quantitative relationship between crack size and capsule size, capsule concentration in capsule-based self-healing virtual cementitious material. Moreover, an analytical expression of probability of a crack penetrating with randomly dispersed capsules is developed in two-dimensional material matrix setup. The influences of the induced rupturing modes of capsules embedded on the self-healing efficiency are analyzed. Much attention is paid to compare the penetrating probability and the hitting probability, in order to assist the designer to make a choice of the optimal rupturing modes of capsules embedded. The accuracy of results of the theoretical model is also compared with Monte-Carlo numerical analysis of crack interacting with capsules. It shows that the developed probability characteristics of a crack interaction with capsules for different rupturing modes is helpful to provide guidelines for designer working with capsule-based self-healing cementitious materials. DOI: http://dx.doi.org/10.5755/j01.ms.23.3.16888

Effects of skim milk concentrate dry matter and spray drying air temperature on formation of capsules with varying particle size and the survival microbial cultures in a microcapsule matrix

ABSTRACTThe impact of total solid (TS) content in combination with the feed rate and air inlet temperature on the survival of Lactobacillus paracasei ssp. paracasei F19 after spray drying in a skim milk matrix has been investigated and correlated with the capsule size. Depending on the experimental conditions, the survival rates ranged from 64 to 0.2%. The higher the air inlet temperature, the lower was the survival rate and an inversely correlation between the TS content and particle size has been determined. These results clearly indicate that process stress analyses and product-related characteristics must not be regarded separately.

Correlation between investment in sexual traits and valve sexual dimorphism in Cyprideis species (Ostracoda).

Assessing the long-term macroevolutionary consequences of sexual selection has been hampered by the difficulty of studying this process in the fossil record. Cytheroid ostracodes offer an excellent system to explore sexual selection in the fossil record because their readily fossilized carapaces are sexually dimorphic. Specifically, males are relatively more elongate than females in this superfamily. This sexual shape difference is thought to arise so that males carapaces can accommodate their very large copulatory apparatus, which can account for up to one-third of body volume. Here we test this widely held explanation for sexual dimorphism in cytheroid ostracodes by correlating investment in male genitalia, a trait in which sexual selection is seen as the main evolutionary driver, with sexual dimorphism of carapace in the genus Cyprideis. We analyzed specimens collected in the field (C. salebrosa, USA; C. torosa, UK) and from collections of the National Museum of Natural History, Washington, DC (C. mexicana). We digitized valve outlines in lateral view to obtain measures of size (valve area) and shape (elongation, measured as length to height ratio), and obtained several dimensions from two components of the hemipenis: the muscular basal capsule, which functions as a sperm pump, and the section that includes the intromittent organ (terminal extension). In addition to the assessment of this primary sexual trait, we also quantified two dimensions of the male secondary sexual trait—where the transformed right walking leg functions as a clasping organ during mating. We also measured linear dimensions from four limbs as indicators of overall (soft-part) body size, and assessed allometry of the soft anatomy. We observed significant correlations in males between valve size, but not elongation, and distinct structural parts of the hemipenis, even after accounting for their shared correlation with overall body size. We also found weak but significant positive correlation between valve elongation and the degree of sexual dimorphism of the walking leg, but only in C. torosa. The correlation between the hemipenis parts, especially basal capsule size and male valve size dimorphism suggests that sexual selection on sperm size, quantity, and/or efficiency of transfer may drive sexual size dimorphism in these species, although we cannot exclude other aspects of sexual and natural selection.

Counts of MSC in the Bone Marrow of Young and Old CBA Mice after a Single Exposure to Osteogenic Stimuli (Curettage, BMP-2 Injection) or Antigens (S. typhimurium Antigenic Complex) and in Heterotopic Bone Marrow Transplants

The efficiency of cloning and the content of multipotent stromal cells (MSC) in the femoral bone marrow of intact CBA mice was 1.5 times less in old mice (24-36 months) than in young ones (2-3 months). The concentration of osteogenic MSC was higher in old vs. young mice (42±3 vs. 22±2%, respectively). Changes in the total counts of MSC and concentrations of osteogenic MSC in response to osteogenic (curettage, BMP-2) and immunogenic stimuli (S. typhimurium antigenic complex) were similar in young and old mice in comparison with intact controls of respective age. The counts of the total pool of bone marrow MSC and pool of osteogenic MSC in response to osteogenic stimuli were 1.5-2 times less in old vs. young mice. This difference seemed to be a result of age-specific decrease of their bone marrow count but not of age-specific decrease of the MSC functional activity, this leading to a decrease in the transplantability of bone marrow stromal tissue of old mice. Comparison of transplantations "old donor - young recipient" vs. "young donor - young recipient" demonstrated a decrease in the count of nuclear cells (1.8 times), size of bone capsule (2-fold), efficiency of MSC cloning (1.6 times), count of MSC per transplant (2.9 times), and count of osteogenic MSC per transplant (3.3 times). The concentrations of osteogenic MSC in transplants from young and old donors leveled in young recipients, that is, seemed to be regulated by the host. Serum concentrations of IL-10 and TNF-α in intact old mice were at least 2.9 and 2 times higher than in young animals, while the concentrations of almost all the rest studied cytokines (IL-2, IL-5, GM-CSF, IFN-γ, IL-4, IL-12) were lower. Presumably, the decrease in the content of bone marrow MSC and in transplantability of bone marrow stromal tissue in old mice were caused by exhaustion of the MSC pool as a result of age-specific chronic inflammation. These data indicated a close relationship between age-specific changes in the stromal tissue and immune system.

Modeling and optimization of the parameters affecting the in-situ microencapsulation process for producing epoxy-based self-healing anti-corrosion coatings

Micro/nanocapsules of urea–formaldehyde resin loaded with linseed oil, which are a self-healing agent in glass flake epoxy anti-corrosion paint, were prepared using a combination of ultrasonic homogenization and in-situ polymerization. The main objective of this study was to model and optimize the microencapsulation process. Five-level central composite design was used to design, model, and optimize the microencapsulation process. A quadratic model was constructed to show the dependency of the percentage of encapsulated linseed oil and capsule size, as model responses, on the studied independent variables (the rotational speed of the agitator and the power and duration of sonication). Analysis of variance showed that all of the variables have significant effects on the encapsulated linseed oil percentage, while the rotational speed of the agitator and sonication time is effective variables for controlling the capsule size. Under the determined optimum conditions, a maximum encapsulated linseed oil percentage (ELO%) of 93.9% and a minimum micro/nanocapsule size of 0.574μm were achieved at 594rpm agitation, 350W sonication power, and 3min sonication time. Validation of the model was performed. The percentage relative errors between the predicted and experimental values of the ELO% and micro/nanocapsule size are 1.28% and 3.66%, respectively. The efficacy of the optimum micro/nanocapsules in healing cracks in a glass flake epoxy paint and corrosion protection was investigated by the salt spray test and Tafel polarization technique.

Dietary supplementation with n-3-fatty acids in patients with pancreatic cancer and cachexia: marine phospholipids versus fish oil - a randomized controlled double-blind trial

BackgroundLike many other cancer patients, most pancreatic carcinoma patients suffer from severe weight loss. As shown in numerous studies with fish oil (FO) supplementation, a minimum daily intake of 1.5 g n-3-fatty acids (n-3-FA) contributes to weight stabilization and improvement of quality of life (QoL) of cancer patients. Given n-3-FA not as triglycerides (FO), but mainly bound to marine phospholipids (MPL), weight stabilization and improvement of QoL has already been seen at much lower doses of n-3-FA (0,3 g), and MPL were much better tolerated. The objective of this double-blind randomized controlled trial was to compare low dose MPL and FO formulations, which had the same n-3-FA amount and composition, on weight and appetite stabilization, global health enhancement (QoL), and plasma FA-profiles in patients suffering from pancreatic cancer.MethodsSixty pancreatic cancer patients were included into the study and randomized to take either FO- or MPL supplementation. Patients were treated with 0.3 g of n-3-fatty acids per day over six weeks. Since the n-3-FA content of FO is usually higher than that of MPL, FO was diluted with 40% of medium chain triglycerides (MCT) to achieve the same capsule size in both intervention groups and therefore assure blinding. Routine blood parameters, lipid profiles, body weight, and appetite were measured before and after intervention. Patient compliance was assessed through a patient diary. Quality of life and nutritional habits were assessed with validated questionnaires (EORTC-QLQ-C30, PAN26). Thirty one patients finalized the study protocol and were analyzed (per-protocol-analysis).ResultsIntervention with low dose n-3-FAs, either as FO or MPL supplementation, resulted in similar and promising weight and appetite stabilization in pancreatic cancer patients. MPL capsules were slightly better tolerated and showed fewer side effects, when compared to FO supplementation.ConclusionThe similar effects between both interventions were unexpected but reliable, since the MPL and FO formulations caused identical increases of n-3-FAs in plasma lipids of included patients after supplementation. The effects of FO with very low n-3-FA content might be explained by the addition of MCT. The results of this study suggest the need for further investigations of marine phospholipids for the improvement of QoL of cancer patients, optionally in combination with MCT.

Novel Antifungal Activity for the Lectin Scytovirin: Inhibition of Cryptococcus neoformans and Cryptococcus gattii.

Pathogenic cryptococci are encapsulated yeast that can cause severe meningoencephalitis. Existing therapeutic options are dated and there is a growing need for new alternative antifungal agents for these fungi. Here we report novel inhibition of pathogenic cryptococci by the antimicrobial lectin Scytovirin. Inhibition was most potent against Cryptococcus neoformans var neoformans and C. gattii, with MFC values of 500 nM. Scytovirin binding was localized to the cell wall and shown to affect capsule size and release. No effect was observed on melanization or with cells grown in the presence the cell wall stressor Congo red. Synergy with existing antifungals was indicated, most strongly for amphotericin B. Overall, Scytovirin serves as a much needed new avenue for anticryptococcal development.

Linaria semnanensis (Plantaginaceae), a new species from Iran

Linaria semnanensis is described as a new species discovered in Semnan Province. This newly discovered species is compared with its closest relatives L. grandiflora and L. dalmatica. The new species is similar to L. grandiflora in bract size, stem height and seed form and differs in leaf width, capsule size, adaxial lip sinus of corolla, style length, the length of testa cell and ornamentation of the seed coat. L. semnanensis is similar to L. dalmatica in capsule size, seed shape and size and differs in corolla, spur, pedicel and calyx lobe size, stem height, length of adaxial lip sinus of corolla, distance between apex of adaxial lips of corolla, testa cells shape and ornamentation of the seed coat. Information about the species’ morphology, micromorphology, habitats and distribution is provided.

Sealed accelerants facilitate epoxy asphalt concretes opening to traffic quickly

Epoxy asphalt composites (EACs) have been recommended to be a prior choice to obtain long-life pavements. However, the present anhydrides-cured EACs have to be field-maintained in high summer for over 45days to achieve their ultimate performance. To be opened to traffic immediately, sealed accelerants, triggered by the compact pressure when paved, were employed to accelerate post-cure of EACs. Compared with microcapsules complicatedly synthesized by interfacial polymerization, sealed macroporous-resins containing accelerants, which was simply prepared by covering accelerant-soaked macroporous-resins with polytetrafluoroethylene (PTFE), made it possible to be opened to traffic quickly. It is attributed to the bigger capsule sizes and higher encapsulation efficiencies for them to be easier crushed by aggregates and encapsulate enough accelerants to be released when compacted. This paper reports the facile, low-cost and scalable approach to make anhydrides-cured EACs open to traffic quickly which facilitates EACs to be ideal long-life paving materials.

PromotingLactobacillus caseiandBifidobacterium adolescentissurvival by microencapsulation with different starches and chitosan and poly L-lysine coatings in ice cream

In this research, microencapsulation of the probiotics Lactobacillus casei ATCC 39392 and Bifidobacterium adolescentis ATCC 15703 was performed using calcium alginate, wheat, rice, and high-amylose corn (Hylon VII) starches along with chitosan and poly L-lysine coatings. The effect of microencapsulation on the survival and sensory properties of ice cream over 100 days at −30 °C was evaluated. Scanning electron and optical microscopy were employed to measure capsule size and morphology. The results suggested that the survival of probiotics is increased by microencapsulation. Coating the capsules with chitosan and poly L-lysine led to enhanced bacterial viability and an increase in the size of microcapsules. Among different starches, Hylon starch enhanced the survival of probiotics at low temperatures the most. Furthermore, the addition of probiotics in free and encapsulated states did not have a significant effect on the sensory properties, or pH levels of the final product during storage (p > .05). Practical applications Microencapsulation using various hydrocolloids is a commonly used method for enhancing probiotic survival in ice cream during the frozen storage. This study indicates that the microencapsulation of probiotics can enhance probiotic survival in ice cream after 100 days of storage at −30 °C. Chitosan and poly L-lysine coatings significantly improved the survival of encapsulated probiotics during the storage of ice cream. This improvement is attributed to the role of Hylon starch in creating more integrated microcapsule structure. Moreover, sensory evaluation of ice cream revealed that inoculation with the probiotic culture, in either the encapsulated or the free-state, had no significant effect on texture, color, flavor, taste, or general sensory characterization of ice cream during the storage period at −30 °C (p > .05).

Rapid preparation of polysaccharide hydrogel capsule free of organic reagents to control drug release

Polysaccharide-based hydrogels for drug delivery systems have attracted considerable research. The less use of cross-linking agents is beneficial in reducing the side effects on humans. The aim of this study is to prepare a drug delivery vehicle which allows the drug to be less likely or not released in the stomach but is usually released in the intestine. Here, we synthesized hydrogel capsules with cellulose as carrier and sodium alginate (SA) as shell, respectively. The hydrogel capsule break-up process was controlled by varying the concentration of CaCl2 and SA, the weight ratio of sodium carboxymethylcellulose (CMC) and hydroxyethyl cellulose (HEC) and the size of hydrogel capsules. In addition, the hydrogel capsules showed controlled release under simulated gastric acid conditions by continuous gastrointestinal simulations and showed sustained release in simulated intestinal fluid.

Transcriptomic and virulence factors analyses of Cryptococcus neoformans hypoxia response.

Cryptococcus neoformans is an environmental pathogen requiring atmospheric levels of oxygen for optimal growth. Upon inhalation, C.neoformans disseminates to the brain and causes meningoencephalitis. However, the mechanisms by which the pathogen adapts to the low-oxygen environment in the brain have not been investigated. We isolated a C.neoformans strain with a small capsule from a host tissue, although this strain produces large capsules in normoxic conditions. We hypothesize that this difference in capsule size is attributed to hypoxia caused by chronic inflammatory response. This study investigated the effect of hypoxia on virulence factors (including capsule, melanin, urease, and phospholipase) of C.neoformans and conducted transcriptomic analyses of the virulence-associated genes. We found that C.neoformans grew under hypoxic condition, albeit slowly, and that hypoxia may have inhibited the capsule size, melanin production, and phospholipase and urease activities in C.neoformans.

Physiological Differences in Cryptococcus neoformans Strains In Vitro versus In Vivo and Their Effects on Antifungal Susceptibility.

Cryptococcus neoformans is an environmentally ubiquitous fungal pathogen that primarily causes disease in people with compromised immune systems, particularly those with advanced AIDS. There are estimated to be almost 1 million cases per year of cryptococcal meningitis in patients infected with human immunodeficiency virus, leading to over 600,000 annual deaths, with a particular burden in sub-Saharan Africa. Amphotericin B (AMB) and fluconazole (FLC) are key components of cryptococcal meningitis treatment: AMB is used for induction, and FLC is for consolidation, maintenance and, for occasional individuals, prophylaxis. However, the results of standard antifungal susceptibility testing (AFST) for AMB and FLC do not correlate well with therapeutic outcomes and, consequently, no clinical breakpoints have been established. While a number of explanations for this absence of correlation have been proffered, one potential reason that has not been adequately explored is the possibility that the physiological differences between the in vivo infection environment and the in vitro AFST environment lead to disparate drug susceptibilities. These susceptibility-influencing factors include melanization, which does not occur during AFST, the size of the polysaccharide capsule, which is larger in infecting cells than in those grown under normal laboratory conditions, and the presence of large polyploid "titan cells," which rarely occur under laboratory conditions. Understanding whether and how C. neoformans differentially expresses mechanisms of resistance to AMB and FLC in the AFST environment compared to the in vivo environment could enhance our ability to interpret AFST results and possibly lead to the development of more applicable testing methods.

Enzymatic characterization of clinical and environmental Cryptococcus neoformans strains isolated in Italy

BackgroundCryptococcus neoformans is an encapsulated yeast causing mainly opportunistic infections. The virulence factors involved in cryptococcosis pathogenesis include the presence and the size of the polysaccharide capsule, the production of melanin by phenoloxidase, the growth at 37°C and the enzyme secretion like proteinase, phospholipase and urease. Many other enzymes are secreted by C. neoformans but their role in the fungus virulence is not yet known. AimsIn order to investigate this topic, we compared the phospholipase production between strains from patients and from bird droppings, and we examined its relationship to phenoloxidase production. We further characterized the strains by determining the activity of 19 different extracellular enzymes. MethodsTwo hundred and five Italian C. neoformans clinical isolates and 32 environmental isolates were tested. Phenoloxidase production was determined by the development of brown colonies on Staib's agar. Extracellular phospholipase activity was performed using the semiquantitative egg-yolk plate method. API ZYM commercial kit was used to observe the production and the activity of 19 different extracellular enzymes. ResultsStatistical analysis of the results showed a significantly higher phospholipase activity in the clinical isolates than in the environmental isolates. No significant difference about the phenoloxidase production between both groups was found. Regarding the 19 extracellular enzymes tested using the API ZYM commercial kit, acid phosphatase showed the highest enzymatic activity in both groups. Concerning the enzyme α-glucosidase, the clinical isolates presented a significantly higher positivity percentage than the environmental isolates. A hundred percent positivity in the enzyme leucine arylamidase production was observed in both groups, but the clinical isolates metabolized a significantly greater amount of substrate. ConclusionsThe higher phospholipase production in the clinical isolates group confirms the possible role of this enzyme in the cryptococcosis pathogenesis. The extracellular activities of the enzymes acid phosphatase, α-glucosidase and leucine arylamidase, tested by means of the API ZYM commercial kit, appear to be very interesting. Many studies indicate that these enzymes are involved in the virulence of bacteria and parasites; our results suggest their possible role as virulence factors in Cryptococcus infections too.

Stimuli-Responsive Capsule Membranes for Controlled Release in Pharmaceutical Applications.

In conventional drug delivery, the drug concentration in the blood raises once the drug taken, and then peaks and declines. Since each drug has a level above which it is toxic and another level below which it is ineffective, the drug concentration in a patient at a particular time depends on compliance with the prescribed routine. To achieve more effective efficacy and fewer side effects of drugs, the drug carriers with desirable dosing and controllable release property of drugs are highly desired. Stimuli-responsive capsules with smart gating membranes or hydrogel-based membranes as capsule shells are ideal candidates. The smart capsule membranes enable efficient encapsulation of drugs within the large inner volume, and the responsive gating membranes or hydrogel-based membranes could control the release rate of encapsulated drugs in responding to environmental stimuli. The trigger stimuli could be either artificial or natural ones corresponding to specific diseases, such as temperature, pH, glucose concentration, specific ion, light, and magnetic field. This review highlights the recent development in stimuli-responsive capsule membranes for controlled release in pharmaceutical applications, including two types of stimuli-responsive capsule membranes with different architectures for on/off release and burst release, which can achieve potential uses of case-dependent on/off release and burst release. The preponderances of the smart capsule membranes are that the capsules are with controllable inner space for drug vehicles with desired dose and stimuli-responsive membrane as shell to release drugs at a desired site and/or moment. However, the actual difficulties for the stimuli-responsive capsule membrane systems to go before they can be applied widely in the biomedical fields are discussed. The future works should focus on the improvements of biocompatibility, biodegradability and stimuli-responsiveness of the capsule membranes, easy and scalable fabrication techniques with further decrease of the capsule size for more efficient in vivo applications, and the diversification of the multi-compartmental capsule architectures with multi-stimuli-responsive characteristics for controlled release.