Methodological Considerations on the Use of Different Spectral Decomposition Algorithms to Study Hippocampal Rhythms.

Local field potential (LFP) oscillations are primarily shaped by the superposition of postsynaptic currents. Hippocampal LFP oscillations in the 25- to 50-Hz range ("slow γ") are proposed to support memory retrieval independent of other frequencies. However, θ harmonics extend up to 48 Hz, necessitating a study to determine whether these oscillations are fundamentally the same. We compared the spectral analysis methods of wavelet, ensemble empirical-mode decomposition (EEMD), and Fourier transform. EEMD, as previously applied, failed to account for the θ harmonics. Depending on analytical parameters selected, wavelet may convolve over high-order θ harmonics due to the variable time-frequency atoms, creating the appearance of a broad 25- to 50-Hz rhythm. As an illustration of this issue, wavelet and EEMD depicted slow γ in a synthetic dataset that only contained θ and its harmonics. Oscillatory transience cannot explain the difference in approaches as Fourier decomposition identifies ripples triggered to epochs of high-power, 120- to 250-Hz events. When Fourier is applied to high power, 25- to 50-Hz events, only θ harmonics are resolved. This analysis challenges the identification of the slow γ rhythm as a unique fundamental hippocampal oscillation. While there may be instances in which slow γ is present in the rat hippocampus, the analysis presented here shows that unless care is exerted in the application of EEMD and wavelet techniques, the results may be misleading, in this case misrepresenting θ harmonics. Moreover, it is necessary to reconsider the characteristics that define a fundamental hippocampal oscillation as well as theories based on multiple independent γ bands.

Theta-gamma cascades and running speed.

Oscillations in the hippocampal local field potential at theta and gamma frequencies are prominent during awake behavior and have demonstrated several behavioral correlates. Both oscillations have been observed to increase in amplitude and frequency as a function of running speed. Previous investigations, however, have examined the relationship between speed and each of these oscillation bands separately. Based on energy cascade models where "…perturbations of slow frequencies cause a cascade of energy dissipation at all frequency scales" (Buzsaki G. Rhythms of the Brain, 2006), we hypothesized that cross-frequency interactions between theta and gamma should increase as a function of speed. We examined these relationships across multiple layers of the CA1 subregion, which correspond to synaptic zones receiving different afferents. Across layers, we found a reliable correlation between the power of theta and the power of gamma, indicative of an amplitude-amplitude relationship. Moreover, there was an increase in the coherence between the power of gamma and the phase of theta, demonstrating increased phase-amplitude coupling with speed. Finally, at higher velocities, phase entrainment between theta and gamma increases. These results have important implications and provide new insights regarding how theta and gamma are integrated for neuronal circuit dynamics, with coupling strength determined by the excitatory drive within the hippocampus. Specifically, rather than arguing that different frequencies can be attributed to different psychological processes, we contend that cognitive processes occur across multiple frequency bands simultaneously with organization occurring as a function of the amount of energy iteratively propagated through the brain. NEW & NOTEWORTHY Often, the theta and gamma oscillations in the hippocampus have been believed to be a consequence of two marginally overlapping phenomena. This perspective, however, runs counter to an alternative hypothesis in which a slow-frequency, high-amplitude oscillation provides energy that cascades into higher frequency, lower amplitude oscillations. We found that as running speed increases, all measures of cross-frequency theta-gamma coupling intensify, providing evidence in favor of the energy cascade hypothesis.

Engineered channels enhance cellular density in perfused scaffolds.

Scaffold-based tissue engineering provides cells with an engineered matrix to enhance and direct cell attachment, proliferation and differentiation. One critical limitation to current tissue engineering approaches is the inability to create densely populated constructs thicker than a few 100 µm. We hypothesized that development of porous, channeled scaffolds would increase cell density and uniformity of their spatial distribution through scaffold channel perfusion. Patterned polyurethane sheets were fabricated using a sprayed phase separation technique and laminated together to form 1.5 mm thick channeled scaffolds. Hydraulic permeability testing confirmed the presence of functional channels throughout the multilaminate construct. A continuous flow bioreactor was used to perfuse the construct with medium during the culture period. Cross-sectional cell densities and spatial uniformities were measured in channeled and nonchanneled scaffolds under different seeding and culture conditions. Channeled scaffolds were found to have higher densities of human mesenchymal stem cells than nonchanneled samples. Perfused scaffolds had more uniform spatial distribution of cells within the scaffold compared to statically cultured scaffolds. In conclusion, we have shown the channeled scaffolds to be a promising approach toward creating thick tissue-engineered constructs.

Regular debridement is the main tool for maintaining a healthy wound bed in most chronic wounds.

Sharp debridement is the most clinically and cost-effective way of physically removing and suppressing a biofilm. Continued debridement, as part of a multifaceted treatment strategy, will keep the biofilm in a weakened state.

Effects of biofilm treatments on the multi-species Lubbock chronic wound biofilm model.

OBJECTIVE: To evaluate the efficacy of several biofilm effectors in inhibiting biofilm formation in an in vitro multi-species chronic wound biofilm model. METHOD: The Lubbock Chronic Wound Biofilm (LCWB) model has been described in detail elsewhere. Pathogens used in the model are Pseudomonas aeruginosa, Enterococcus faecalis and Staphylococcus aureus. These are three of the most important species associated with biofilms. Here, the model was exposed to the following biofilm effectors: xylitol, salicylic acid, farnesol, erythritol and two proprietary, semi-solid, wound-dressing formulations currently under development (Sanguitec gels). RESULTS: Biofilm formation was completely inhibited in the LCWB model following treatment with 20% xylitol, 10% erythritol, 1,000 microg/ml farnesol, 20mM salicylic acid or 0.1% of either of the two Sanguitec gel formulations. Salicylic acid specifically inhibited S. aureus (p<0.01) at 10mM and 20mM, consequently increasing the ratios of P. aeruginosa and E. faecalis within the biofilm. Xylitol had an increasing inhibitory effect on P. aeruginosa (p<0.01) at all concentrations evaluated. Erythritol had an inhibitory effect on P. aeruginosa and S. aureus growth (p<0.01) at over 5% concentrations. The inhibitory effect of both Sanguitec gel formulations was more broadly effective, with an increasingly inhibitory effect on all LCWB species (p<0.01). CONCLUSION: The LCWB model provides a multi-species format with which to evaluate the effect of biofilm effectors on wound flora in a biofilm phenotype. These results suggest that different treatments can target specific populations within a biofilm. Salicylic acid preferentially targeted S. aureus, xylitol preferentially targeted P. aeruginosa, while erythritol preferentially targeted both P. aeruginosa and S. aureus. In contrast, the two Sanguitec gel formulations provided a broad, less preferential, inhibition of biofilm development. DECLARATION OF INTEREST: Research and Testing Laboratory is a for-profit enterprise that develops molecular methods and performs service research work on biofilms. Sanguitec gel was developed by JPK and CEJ.

Characterization and performance of membranes designed for macroencapsulation/implantation of pancreatic islet cells.

Amphiphilic polymer membranes were synthesized for macroencapsulation of cells and characterized by select chemical and biological techniques. The membranes were prepared by crosslinking hydrophilic poly(N,N-dimethyl acrylamide) (PDMAAm) main chains with hydrophobic di-, tri-, and octa-methacrylate telechelic polyisobutylene (PIB) stars. The hydrophilic/hydrophobic composition and the molecular weights between crosslink sites (both M(c,hydrophilic) and M(c,hydrophobic)) were controlled by synthesis conditions. Small tubular membranes were made by in situ rotational copolymerization/crosslinking and filled with pancreatic rat islets. The water-swelling behavior, mechanical properties, and oxygen and insulin diffusion were studied. Macroencapsulatory performance of these membranes was investigated in vitro by macroencapsulation of pancreatic rat islets within tubular membranes for up to 1.5 months, and studying the insulin secreting ability of encapsulated islets in culture. The membranes are robust and maintain their integrity for the period of encapsulation. They allow oxygen and insulin diffusion. Macroencapsulated islets maintained their viability and insulin secretion over an extended period (i.e., 45 days).

Synthesis, permeability and biocompatibility of tricomponent membranes containing polyethylene glycol, polydimethylsiloxane and polypentamethylcyclopentasiloxane domains.

The synthesis of "smart" tricomponent amphiphilic membranes containing poly(ethylene glycol) (PEG), polydimethylsiloxane (PDMS) and polypentamethylcyclopentasiloxane (PD(5)) domains is described. Contact angle hysteresis indicates that in air, the surfaces of such PEG/PD(5)/PDMS membranes are enriched by the hydrophobic components, PDMS and PD(5), while in water, the surfaces are rich in the hydrophilic PEG. The oxygen permeability of a series of membranes with varying M(c,hydrophilic) (M(n,PEG)=4600, 10,000 and 20,000 g/mol) and varying PEG/PD(5)/PDMS compositions was studied. Oxygen permeability increased with the amount of PDMS in the membrane. The molecular weight cut-off (MWCO) ranges and permeability coefficients of insulin through a series of PEG/PD(5)/PDMS(=29/14/57) membranes with varying M(c,hydrophilic) were determined. Insulin permeability is directly related to M(c,hydrophilic) of the membrane. MWCO studies show that the membranes are semipermeable to, i.e., allow the transport of smaller proteins such as insulin (M(n)=5733 g/mol, R(s)=1.34 nm) and cytochrome c (M(n)=12,400 g/mol, R(s)=1.63 nm), but are barriers to larger proteins such as albumin (M(n)=66,000 g/mol, R(s)=3.62 nm). Implantation of representative membranes in rats showed them to be biocompatible. According to these studies, PEG/PD(5)/PDMS membranes may be suitable for biological applications, e.g., immunoisolation of cells.

Intellectual ability in adolescents with conduct disorder.

A U.K. population of young offenders show the typical deficits in Verbal IQ reported in conduct disordered populations. Performance IQs are generally lower than those of previous studies from the 1970s, raising a question about decline in intellectual ability.

Novel polyisobutylene/polydimethylsiloxane bicomponent networks: III. Tissue compatibility.

The tissue biocompatibility of a series of novel rubbery polyisobutylene (PIB)/polydimethylsiloxane (PDMS) bicomponent networks was investigated by in vivo implantation into rats. Bicomponent networks of varying composition (PIB wt%/PDMS wt% = 70/30, 50/50, 35/65) as well as a standard polyethylene control were implanted intraperitoneally. After eight weeks the implants and surrounding tissue were removed for histological evaluation. In all scoring categories (i.e. collagen thickness, fibrous tissue orientation, collagen deposition in muscle tissue, lymphocyte infiltration, angiogenesis) the PIB/PDMS bicomponent network implants elicited either less or similar tissue and cellular response than polyethylene. To determine which implant elicited the least tissue and cellular response overall, a weighted score including collagen thickness, lymphocyte infiltration, and angiogenesis was calculated for each implant. According to these preliminary investigations, PIB/PDMS bicomponent networks are suitable for implant applications.

Evaluation of extended-release applications for solid dispersion hot-melt fluid bed coatings utilizing hydrophobic coating agents.

A new hot-melt fluid bed coating method was evaluated for potential extended-release applications. Chlorpheniramine maleate (CPM) USP was chosen as a model drug. The assays for drug release and content uniformity were dictated by the USP Official Monograph for a Chlorpheniramine Maleate Extended-Release Capsule. The fluid bed chamber was charged with CPM-loaded nonpareils and hydrophobic coating agents in the solid state. The method consists of four processing stages: (a) warming, (b) preheating, (c) melting-spreading, and (d) cooling-congealing. Various hydrophobic coating agent candidates were evaluated for extended-release potential by a preliminary screen at a coating agent level of 1.5% (w/w). A beeswax coating agent was identified as the most promising candidate of the preliminary screen. After the level of beeswax was increased to 2.0%, the dissolution profile met all of the specifications of the USP Drug Release Test 1 for a CPM Extended-Release Capsule. The potency and content uniformity remained unchanged by the process. Dual coatings demonstrated a cumulative extension of release superior to the capability of a single coat. The new method is a viable alternative to hot-melt spray-coating methodologies. Organic solvents, spraying equipment, steam jackets, and/or heating tape are eliminated from the process. A reduction of equipment costs, setup time, and cleanup time may be realized. The method has demonstrated extended-release capabilities. No excessive attrition of potency or content uniformity has been noted. Additive, multiple coatings that have a cumulative effect on release retardation are feasible.

Preliminary assessment of an image analysis method for the evaluation of pharmaceutical coatings.

This article describes the development and preliminary assessment of an innovative image analysis method for the evaluation of pharmaceutical coatings. Nonpareils previously hot-melt coated with a red water-soluble dye incorporated into polyethylene glycol were employed to develop and evaluate the new method. Digital images of batch samples were acquired and transferred to a PC for evaluation by image analysis software. Standard deviations generated for the optical densities of individual nonpareils in each batch sample were utilized to evaluate coating uniformity. Mean optical densities generated for each batch sample were used to evaluate the mean coating thickness. Method precision determinations, using an average sample size of 221 nonpareils, returned a relative standard deviation of 2.47% for coating uniformity and 1.71% for coating thickness. The new method is rapid, objective, and relatively inexpensive. It proved to be both qualitative and quantitative for coating uniformity, qualitative for coating thickness, and maintained a high degree of precision. The method can be used independently or in conjunction with other methods to eliminate their subjective aspects regarding sample selection.

Amphiphilic networks. X. Diffusion of glucose and insulin (and nondiffusion of albumin) through amphiphilic membranes.

Select semipermeable amphiphilic membranes have been prepared and their diffusional characteristics for glucose, insulin, and albumin investigated. The membranes were prepared by cast copolymerization of a hydrophilic monomer (i.e., N,N-dimethyl acrylamide, or N,N-dimethylaminoethyl methacrylate) with the hydrophobic crosslinker methacrylate-ditelechelic polyisobutylene. The products have sufficient mechanical properties for the fabrication of swollen membranes, sheets, tubes, etc. Membranes have been identified which allowed the rapid simultaneous countercurrent diffusion of glucose (Mn = 180 Da) and insulin (Mn = 5733 Da) but did not allow albumin (Mn approximately 60,000 Da) to pass. Evidently, the effective molecular weight cutoff point of these membranes is in the 6-60-KDa range.

Development and optimization of a solid dispersion hot-melt fluid bed coating method.

A new hot-melt fluid bed coating process has been developed, characterized, and optimized. Polyethylene glycol served as the model coating agent and was charged with substrate into the fluid bed chamber in the solid state. The processing stages included: (A) warm-up, (B) preheating, (C) melting-spreading, and (D) cooling-congealing. A central composite design was utilized to characterize and optimize the process. Substrate porosity and density evaluations were conducted by mercury intrusion. The method proved capable of coating nonpareils from 10 to 35 mesh (0.500 to 2.00 mm) and tablets up to 1 g. The nonpareils were coated as individual particles, while particle sizes significantly smaller than 40 mesh (0.420 mm) tended to agglomerate. The porosity and density values of dissimilar nonpareil batches showed a large degree of variation, affecting the method's reproducibility. Additive coatings were achieved by sequential runs using coating agents of diminishing melting points. The method is a viable alternative to hot-melt spray-coating processes. Organic solvents, spraying equipment, steam jackets, and/or heating tape are eliminated from the process.

Polyisobutylene-toughened poly(methyl methacrylate): III. PMMA-l-PIB networks as bone cements.

A series of novel polyisobutylene (PIB)-toughened poly(methyl methacrylate) (PMMA) networks consist of rubbery PIB domains covalently bonded to a glassy PMMA matrix. Materials containing 8.5-17 wt.% PIB (Mn = 18,000 g/mol) in a PMMA matrix (PIB/PMMA) were evaluated to assess their feasibility as the powder component along with methyl methacrylate (MMA) as the liquid component in a standard powder/liquid bone cement formulation. A standard ISO four-point bend test, commonly used for testing bone cements, was employed to investigate flexural properties. The mixing time and powder/liquid (P/L) ratio were studied to formulate novel PIB/PMMA cements for optimum toughness. Appropriate formulations led to improved toughness while adequate flexural strength and modulus were maintained. An experimental PIB/PMMA-system exhibited approximately 57 MPa flexural strength and approximately 2000 MPa flexural modulus.

A comparison of two insecticidal shampoos in the treatment of head louse infection.

The efficacy and tolerability of a phenothrin liquid shampoo was compared with a carbaryl shampoo in 50 children with head lice infection. Twenty-five children were treated with a phenothrin liquid shampoo and 25 with a carbaryl shampoo. Each treatment was applied on three occasions at three-day intervals. Reinspection two weeks after initiation of treatment revealed two apparent treatment failures in the carbaryl group and one in the phenothrin group. No side effects were reported for either treatment. It is concluded that phenothrin liquid shampoo is a safe and effective treatment for head louse infection and is therefore a useful addition to those insecticides currently employed.

Avascular necrosis complicating fracture of the distal tibial epiphysis.

We have described a patient with a very rare case of distal tibial epiphyseal AVN after a severe fracture. This patient subsequently had complete asymptomatic revascularization and closure of the epiphyseal plate without deformity. Practitioners should be aware, however, that in younger persons a similar lesion can lead to premature closure of the plate and resultant deformity.