Photomechanical wave-assisted molecular delivery in oral biofilms

Photomechanical waves (PW), the product of an intense light beam interaction with a target material, enhance molecular delivery across biological membranes and skin. The ability to deliver methylene blue (MB), a fluorescent probe and photosensitizer, into bacterial biofilms was demonstrated by applying PW on saliva-derived multi-species biofilms that were developed on agar surfaces in 24-well plates. PW were generated with a Q-switched Nd:YAG laser and were directed into the biofilms in the presence of 25 μg/ml MB. The biofilms were then irradiated with red light at 665 nm. After illumination, adherent bacteria were scraped and spread over the surface of blood agar plates. Survival fractions were calculated by counting bacterial colonies. Microbial analysis was performed via a colony lift method and a DNA checkerboard assay using whole genomic probes to 40 oral microorganisms. Visual analysis by confocal scanning laser microscopy demonstrated that the application of PW enhanced the penetration depth of MB in biofilms. Exposure to MB, PW and light led to a significant reduction of the mean levels of log₁₀ CFU counts compared with the group that received MB and light (P = 0.006). The DNA checkerboard assay showed some benefit from PW-assisted phototargeting in 25 biofilm microorganisms relative to phototreatment alone. Our data provide a basis for further exploration and optimization of PW parameters for complete eradication of microorganisms in oral microcosm biofilms.     

Recovery of antioxidant phenolics from white vinification solid by-products employing water/ethanol mixtures

Solid wastes from white vinification, including grape peels, seeds and stems, were used as raw material for the recovery of antioxidant polyphenols. Extractions were performed using non-toxic media composed of water/ethanol mixtures and hydrochloric, acetic or tartaric acid. Recovery efficiency was assessed by monitoring the antioxidant potency of extracts and several indices related to their polyphenolic composition, including total polyphenol, total flavonoid, total flavanol and condensed tannin (proanthocyanidin) content. Among the by-products tested, seeds were shown to contain exceptional amounts of total polyphenols (13.76 g per 100g dry weight), followed by stems (7.47 g per 100g dry weight) and peels (0.97 g per 100g dry weight). Extracts with the highest antioxidant activity from all by-products were obtained with 57% ethanol. Acidification of this medium with 0.1% HCl improved polyphenol recovery and antiradical activity for stem extracts, but it was unfavourable for seed extraction.     

Combining immunological and molecular data to assess phylogenetic relations of some Greek Podarcis species

Most recent molecular studies revealed the phylogeny of Greek Podarcis species, which for years remained elusive, due to discordant data produced from various chromosomal, complement fixation and protein studies. In this report, we analyzed cellular immune responses of spleen-derived lymphocytes from six allopatric Podarcis species encountered in Greece, by assessing two-way mixed lymphocyte reaction (MLR)-induced proliferation. On the basis of stimulation indices (S.I.) as determined from cultures set up from xenogeneic splenocytes coincubated in pairs, we generated a phylogenetic tree, fully consistent with the phylogenetic relationships of Podarcis as determined by parallel analyses based on partial mitochondrial (mt) DNA sequences. Although the exact mechanisms triggering lymphocyte responses in lizard two-way xenogeneic MLR are not fully understood, our results show the potential use of cell-mediated immune responses as an additional approach to mtDNA analysis, for species delimitation within specific lizard taxa.     

Colloidal stability of carbon nanotubes in an aqueous dispersion of phospholipid

Within the family of nanomaterials, carbon nanotubes (CNTs) have emerged as a new efficient scaffold for studying molecular interactions at interfaces. Poor dispersability of CNTs in any solvent presents a considerable drawback for the development of novel functional composite structures. Previous studies have demonstrated that the solubility of CNTs can be greatly enhanced by employing appropriate surfactants, some of them being biological molecules. In this work, we study the noncovalent wrapping of lipid chains onto the graphitic surface of single-walled material (SWCNTs) by electron microscopy and Raman spectroscopy. Stable and homogenous aqueous suspensions of SWCNTs in the presence of lipids have been prepared, whereas their electrophoretic mobility was confirmed by zeta-potential measurements. Raman measurements revealed that smaller diameter SWCNTs are preferentially dispersed by lipid molecules in the aqueous supernatant part of the prepared suspension.     

The influence of the location of a nest box in an individually ventilated cage on the preference of mice to use it

The improvement of housing conditions for mice by using environmental enrichment materials is of high concern for the scientific community. Plastic, autoclavable nest boxes are commercially available and ready to use for specific cases such as in individually ventilated cages, metabolic cages, or during toxicological studies. The aim of this study was to see if the location of the nest box within the cage could influence the mice to prefer and use it. Located on the cage floor or hung from the cage lid, a nest box (MPLEX, Otto Environmental, Milwaukee, Wisconsin), enriched the cages. The study concluded that the location of the nest boxes in the individually ventilated cage plays a significant role in the mice preferring to use it or to avoid it. It is also important to use environmental enrichment items that provide animals with the possibility of expressing their preferences and manipulating them in a way to cope better with their environmental conditions.     

Immune status evaluation of patients with chronic heart failure

Chronic heart failure (CHF) may be considered a state of immune activation and persistent inflammation expressed by increased circulating levels of pro- and anti-inflammatory cytokines. The purpose of the study was to investigate the immune status in patients with CHF compared to normal individuals. We measured serum cytokine levels as well as cytokine production after ex vivo LPS stimulation of whole blood taken from 14 patients with CHF and 14 healthy volunteers. We used 500 pg/ml of LPS for an incubation period of 4h to stimulate 100 microL of whole blood. Patients with CHF had significantly higher levels of TNF-RI, and TNF-RII in serum compared to normal individuals. TNF-alpha, IL-6, and IL-10 did not differ significantly. After LPS stimulation, patients with CHF had significantly higher levels of TNF-alpha and IL-10, and significantly lower IL-6 levels compared to normal individuals. TNF-alpha receptors did not differ significantly. Patients with CHF may be found in a pro- as well as an anti-inflammatory state. They also do not develop endotoxin tolerance in an ex vivo laboratory model using whole blood stimulated with LPS. They may have increased TNF-alpha and IL-10 production after LPS stimulation of whole blood, which may contribute to a worsening of heart function, more severe disease presentation and a worse outcome during infections.     

A reduction-based exact algorithm for the contact map overlap problem.

Aligning proteins based on their structural similarity is a fundamental problem in molecular biology with applications in many settings, including structure classification, database search, function prediction, and assessment of folding prediction methods. Structural alignment can be done via several methods, including contact map overlap (CMO) maximization that aligns proteins in a way that maximizes the number of common residue contacts. In this paper, we develop a reduction-based exact algorithm for the CMO problem. Our approach solves CMO directly rather than after transformation to other combinatorial optimization problems. We exploit the mathematical structure of the problem in order to develop a number of efficient lower bounding, upper bounding, and reduction schemes. Computational experiments demonstrate that our algorithm runs significantly faster than existing exact algorithms and solves some hard CMO instances that were not solved in the past. In addition, the algorithm produces protein clusters that are in excellent agreement with the SCOP classification. An implementation of our algorithm is accessible as an on-line server at http://eudoxus.scs.uiuc.edu/cmos/cmos.html.     

Secretory structures and cytochemical investigation of the leaf of Phlomis fruticosa, a seasonally dimorphic subshrub. Secreting activity of the leaf-originating calluses

Various species of the genus Phlomis have been reported to produce metabolites demonstrating significant pharmacological efficiency. In mature summer leaves of Phlomis fruticosa several secretory cells and structures were detected. A spectrum of compounds was localized histochemically within them. Calluses produced from leaf-cell cultures were histochemically investigated as well. Callus cells, although of low-differentiation, are synthetically active and produce various compounds. Many of them could be of pharmaceutical interest.     

Morphologic and morphometrical study of the muscle spindle in muscular dystrophy

OBJECTIVE: To determine the morphologic and the morphometrical features of spindles in biopsies of patients with different types of muscular dystrophy and investigate the possible involvement of the spindle in the pathologic process of these diseases. STUDY DESIGN: The following variables were studied in biopsy specimens from 10 patients with Duchenne or Becker dystrophy, 9 with limb-girdle dystrophy, 3 with congenital dystrophy and 3 with facioscapulohumeral dystrophy: diameter and area of spindle; thickness of the capsule; number, diameter and area of intrafusal fibers; and number and area of nuclei. RESULTS: The statistical evaluation of the data showed significant differences regarding the thickness of the capsule, which was greater in patients than in controls, while the diameter and the area of the fibers were all smaller in patients than in controls. The area of nuclei of fibers was increased; this was a common feature for all types of muscular dystrophy. CONCLUSION: These findings indicate that the spindle possibly participates in the pathologic process of different types of muscular dystrophies.     

Temporal progression and extent of the return of sensation in the foot provided by the saphenous nerve after sciatic nerve transection and repair in the rat - implications for nociceptive assessments

Sensory testing, by providing stimuli for nociceptors of the foot, is a popular method of evaluating sensory regeneration after damage to the sciatic nerve in the rat. In the following study, 20 rats were submitted to double transection of the sciatic nerve. The subsequent 14 mm gap was repaired through guidance interponation. In order to evaluate nerve regeneration, sensory testing was performed additionally to other methods, which included motor testing, morphometry, and electron microscopic assessments of nerves. Somatosensory testing revealed that all animals exhibited next to the same amount of sensory reinnervation on their foot regardless of their experimental group. In motor tests, however, two out of the three experimental groups did not improve at all. These groups also failed to show neural regrowth in morphometric and electron microscopic assessments of the associated nerve. Retrograde tracing was able to prove the saphenous nerve as an alternative source of sensory reinnervation in animals with failed sciatic regeneration. This means that results of sensory testing in the rat should be treated with caution, taking into account the areas tested and the likelihood that in these areas saphenous sprouting could have taken place. Furthermore, it is strongly advised that somatosensory testing should be conducted only on toe 5.