Surfactant bilayers maintain transmembrane protein activity.

In vitro studies of membrane proteins are of interest only if their structure and function are significantly preserved. One approach is to insert them into the lipid bilayers of highly viscous cubic phases rendering the insertion and manipulation of proteins difficult. Less viscous lipid sponge phases are sometimes used, but their relatively narrow domain of existence can be easily disrupted by protein insertion. We present here a sponge phase consisting of nonionic surfactant bilayers. Its extended domain of existence and its low viscosity allow easy insertion and manipulation of membrane proteins. We show for the first time, to our knowledge, that transmembrane proteins, such as bacteriorhodopsin, sarcoplasmic reticulum Ca(2+)ATPase (SERCA1a), and its associated enzymes, are fully active in a surfactant phase.

Effect of genotype of growing rabbits on productive performance with special reference to residual feed intake at hot temperature.

OBJECTIVE: Better feed efficiency can be achieved by selecting rabbit genotypes with lower residual feed intake (RFI) under high ambient temperatures. METHODS: Two genotypes of rabbits (Jabali, Saudi local breed and imported, Spanish V-line) were used to derive RFI and to investigate the relationship between RFI and productive traits. In total, 250 animals (125 each) were housed in individual wire mesh cages in a semi-closed rabbitry. Growth performance, feed criteria, carcass evaluation, biochemical blood analysis, and immune responses were determined. RESULTS: Superiority in growth performance, feed efficiency, carcass characteristics, and cellular immunity was recorded in the Jabali breed compared to the V-line genotype. According to regression analysis, a significant effect of daily body weight gain was found, upon computing the expected feed intake in both genotypes. Moreover, mid-body weight0.75 had a significant effect only in the Jabali breed. Positive correlation coefficients between RFI and dry matter feed intake or feed conversion ratio were found. The same trend in this relationship between RFI and productive traits was observed in some cases for both genotypes. An opposite trend in correlations was observed in the studied genotypes for some traits. CONCLUSION: The results suggest that the relationship between RFI and productive traits must be taken into consideration in rabbit breeding programs under the prevailing environment. However, further studies are required to investigate the effect of rabbit genotype and environmental factors on computing RFI.

The Development of Nonthermal Plasma and Tirapazamine as a Novel Combination Therapy to Treat Melanoma In Situ.

Although melanoma accounts for only 5.3% of skin cancer, it results in >75% of skin-cancer-related deaths. To avoid disfiguring surgeries on the head and neck associated with surgical excision, there is a clear unmet need for other strategies to selectively remove cutaneous melanoma lesions. Mohs surgery is the current treatment for cutaneous melanoma lesions and squamous and basal cell carcinoma. While Mohs surgery is an effective way to remove melanomas in situ, normal tissue is also excised to achieve histologically negative margins. This paper describes a novel combination therapy of nonthermal plasma (NTP) which emits a multitude of reactive oxygen species (ROS) and the injection of a pharmaceutical agent. We have shown that the effects of NTP are augmented by the DNA-damaging prodrug, tirapazamine (TPZ), which becomes a free radical only in conditions of hypoxemia, which is often enhanced in the tumor microenvironment. In this study, we demonstrate the efficacy of the combination therapy through experiments with B16-F10 and 1205 Lu metastatic melanoma cells both in vitro and in vivo. We also show the safety parameters of the therapy with no significant effects of the therapy when applied to porcine skin. We show the need for the intratumor delivery of TPZ in combination with the surface treatment of NTP and present a model of a medical device to deliver this combination therapy. The importance of functional gap junctions is indicated as a mechanism to promote the therapeutic effect. Collectively, the data support a novel therapeutic combination to treat melanoma and the development of a medical device to deliver the treatment in situ.

How to best estimate the viscosity of lipid bilayers.

The viscosity of lipid bilayers is a property relevant to biological function, as it affects the diffusion of membrane macromolecules. To determine its value, and hence portray the membrane, various literature-reported techniques lead to significantly different results. Herein we compare the results issuing from two widely used techniques to determine the viscosity of membranes: the Fluorescence Lifetime Imaging Microscopy (FLIM), and Fluorescence Recovery After Photobleaching (FRAP). FLIM relates the time of rotation of a molecular rotor inserted into the membrane to the macroscopic viscosity of a fluid. Whereas FRAP measures molecular diffusion coefficients. This approach is based on a hydrodynamic model connecting the mobility of a membrane inclusion to the viscosity of the membrane. We show that: This article emphasizes the pitfalls to be avoided and the rules to be observed in order to obtain a value of the bilayer viscosity that characterizes the bilayer instead of interactions between the bilayer and the embedded probe.

Effect of genotype and egg weight on hatchability properties and embryonic mortality pattern of native chicken populations.

Fertility and hatchability are 2 major parameters that highly influence the reproductive performance of chicken breeds. The objective of this study is to investigate how the genetic background of chickens affects the aspects of fertility, hatchability, and embryonic mortality pattern. Six different native chicken genotypes (black, black-barred, brown, gray, naked neck, and frizzle) kept under similar conditions were evaluated. A total of 1,645 fertile pedigreed eggs from all genetic groups were collected and incubated in forced draft setter. Fertility, hatchability, embryonic mortality, and hatched chick weight were determined. The data were subjected to a one-way analysis of variance with breed (genotype) as a fixed effect. Sire component of variance were used to compute heritability estimates for hatchability traits. The results showed that the fertility and hatchability of the eggs produced from the naked neck or frizzle genotypes exhibited higher values compared to the other genetic groups. An increase in the relative weight of hatched chicks was detected in hatching eggs weighing 44 g or higher. Therefore, attention should be given to the egg size produced by native chicken populations to achieve maximum hatchability performance. Multiple regression analysis revealed that the settable egg weight and the egg weight loss during incubation are the main factors affecting the relative weight of hatched chicks of all genotypes.

Green Synthesis of Zinc Oxide Nanoparticles Using Red Seaweed for the Elimination of Organic Toxic Dye from an Aqueous Solution.

This study aims to produce green zinc oxide nanoparticles (ZnO-NPs) derived from red seaweed (Pterocladia Capillacea) and evaluate their potential to absorb Ismate violet 2R (IV2R) ions from an aqueous solution. UV-vis spectrophotometry, Fourier-transform infrared spectroscopy (FTIR), scanning electron microscopy (SEM), X-ray diffraction (XRD), and a Brunauer-Emmett-Teller surface area analysis (BET) were used to analyze the structural, morphological, and optical features of the synthesized nanoparticles. The change in color of the chemical solution revealed the formation of zinc oxide nanoparticles. The FTIR examination confirmed the synthesis of both Zn and ZnO nanoparticle powder, with a BET surface area of 113.751 m2 g-1 and an average pore size of 2.527 nm for the synthesized adsorbent. Furthermore, the maximum removal effectiveness of IV2R was 99% when 0.08 g ZnO-NPs was applied at a pH of 6, a temperature of 55 °C, and a contact time of 120 min. The dye adsorption capacity of the ZnO-NPs was 72.24 mg g-1. The adsorption process was also controlled by the Freundlich adsorption model and pseudo-second-order reaction kinetics. The adsorption of IV2R ions onto the ZnO-NPs could be represented as a nonideal and reversible sorption process of a nonuniform surface, according to Freundlich adsorption isotherms. In addition, the constant values of the model parameters were determined using various nonlinear regression error functions. Moreover, thermodynamic parameters such as entropy change, enthalpy change, and free energy change were investigated; the adsorption process was spontaneous and endothermic. The high capacity of the ZnO-NPs synthesized by red seaweed promotes them as promising substances for applications in water treatment for the removal of IV2R dye from aqueous systems.

Physiological Performance, Antioxidant and Immune Status, Columnaris Resistance, and Growth of Nile Tilapia That Received Alchemilla vulgaris-Supplemented Diets.

The current perspective is a pioneering trial to assess the efficacy of the dietary supplementation of Alchemilla vulgaris powder (AVP) in the diet of Nile tilapia (Oreochromis niloticus) on growth performance, blood picture, hepatic and renal biomarkers, immune status, and serum and tissue antioxidant capacity and to investigate the resistance against Flavobacterium columnare challenge. Fish (n = 360) were distributed into six groups (three replicates each) and received increasing AVP supplementation levels (0, 2, 4, 6, 8, and 10 g kg-1) for 60 days. Furthermore, fish were exposed to the bacterial challenge of a virulent F. columnare strain and maintained under observation for 12 days. During the observation period, clinical signs and the cumulative mortality percentage were recorded. The results demonstrated that the growth performance, feed conversion ratio, and hematological profile were noticeably enhanced in the AVP-supplemented groups compared to the control. The most promising results of weight gain and feed conversion ratio were recorded in the groups with 6, 8, and 10 g AVP kg-1 diets in a linear regression trend. The levels of hepatorenal function indicators were maintained in a healthy range in the different dietary AVP-supplemented groups. In a dose-dependent manner, fish fed AVP dietary supplements displayed significant augmented serum levels of innate immune indicators (lysozyme, nitric oxide, and complement 3) and antioxidant biomarkers (Catalase (CAT), superoxide dismutase (SOD), total antioxidant (TAC), and reduced glutathione (GSH) with a marked decrease in myeloperoxidase (MPO) and malondialdehyde (MDA) levels). Likewise, hepatic CAT and SOD activities were significantly improved, and the opposite trend was recorded with hepatic MDA. The highest AVP-supplemented dose (10 g/kg) recorded the highest immune-antioxidant status. Based on the study findings, we highlight the efficacy of AVP as a nutraceutical dietary supplementation for aquaculture to enhance growth, physiological performance, and immune-antioxidant status and as a natural economic antibacterial agent in O. niloticus for sustaining aquaculture. It could be concluded that the dietary supplementation of 10 g AVP/kg enhanced O. niloticus growth, physiological performance, immune-antioxidant status, and resistance against F. columnare.

Recent applications of fluorescence recovery after photobleaching (FRAP) to membrane bio-macromolecules.

This review examines some recent applications of fluorescence recovery after photobleaching (FRAP) to biopolymers, while mainly focusing on membrane protein studies. Initially, we discuss the lateral diffusion of membrane proteins, as measured by FRAP. Then, we talk about the use of FRAP to probe interactions between membrane proteins by obtaining fundamental information such as geometry and stoichiometry of the interacting complex. Afterwards, we discuss some applications of FRAP at the cellular level as well as the level of organisms. We conclude by comparing diffusion coefficients obtained by FRAP and several other alternative methods.

Helix-coil transition of DNA monitored by pressure perturbation calorimetry.

We report the first use of pressure perturbation calorimetry (PPC) to characterize the heat-induced helix-coil transition of DNA polymers. The alternating copolymer poly[d(A-T)] was studied in aqueous solutions containing 5.2 and 18.2 mM Na+; it exhibited helix-coil transition temperatures of 33.6 and 44.7 degrees C, respectively. The transition is accompanied by a negative molar volume change, DeltaV) -2.6 and -2.1 mL/mol (base pair), respectively, and an increase in the coefficient of thermal expansion, Deltaalpha=+5x10(-4) K(-1) (at both ionic strengths). These values are consistent with a greater hydration of the coil form. The larger water-accessible surface area of the coil causes more water molecules to assume a bound, more densely packed structure that then gradually decreases with increasing temperature, leading to a larger value of R. The magnitude of the volume changes detected by PPC were larger than those deduced from high-pressure UV spectroscopy, shedding light on the effect of pressure on DeltaV. The shape of the PPC peak was nearly identical to the shape of the DSC peak, providing direct evidence for the correlation between the molar volume change and enthalpy change for the helix to coil transition of DNA.

Comparison of the heat- and pressure-induced helix-coil transition of two DNA copolymers.

The helix-coil transition of poly[d(I-C)] and poly[d(A-T)] was studied as a function of hydrostatic pressure, temperature, and sodium ion concentration. These studies were undertaken in light of a recently published phase diagram for double stranded nucleic acids [Dubins et al. J. Am. Chem. Soc. 2001, 123, 9254-9259]. The sign and magnitude of the volume change for the heat-induced helix-coil transition, DeltaV(T), of poly[d(I-C)] and poly[d(A-T)] were dependent on the helix-coil transition temperature, T(M), at atmospheric pressure. The sign of DeltaV(T) changed from negative to positive as T(M) was increased by increasing the sodium ion concentration. For poly[d(I-C)], DeltaV(T) = 0 cm(3) mol(-1), when the sodium ion concentration is such that the spectroscopically monitored T(M) = 55 degrees C at atmospheric pressure. For poly[d(A-T)], the value of DeltaV(T) = 0 under conditions such that T(M) = 47 degrees C at atmospheric pressure. Negative values of DeltaV(T) imply that the helical form is destabilized at high pressure. Under experimental conditions where the DeltaV(T) for the transition is negative, the transition could be caused by increasing the pressure under isothermal conditions. At temperatures below T(M) measured at atmospheric pressure the midpoint of the pressure-induced helix-coil transition, P(M), decreases with increasing temperature. The volume change of the pressure-induced transitions helix-coil transition, DeltaV(P), was calculated assuming a two-state model. The magnitude of DeltaV(P) (per cooperative length) was much larger than the volume change (per base pair) measured for the heat-induced transition, DeltaV(T), calculated using the Clapeyron equation. The ratio of these two volume changes was used to calculate the cooperative length for the pressure-induced transition. This parameter depends strongly on temperature, becoming greater closer to T(M) measured at atmospheric pressure. At temperatures approaching T(M) the magnitude of the cooperative length of the pressure-induced transition is approximately twice that observed for the heat-induced transition (N(T)). On the basis of the temperature dependence of the DeltaV(T) for the two polymers the coefficient of thermal expansion of the two polymers was found to be 0.17 and 0.16 cm(3) K(-1) mol(-1) for poly[d(I-C)] and poly[d(A-T)], respectively.

A look at the effect of sequence complexity on pressure destabilisation of DNA polymers.

Our previous studies on the helix-coil transition of double-stranded DNA polymers have demonstrated that molar volume change (ΔV) accompanying the thermally-induced transition can be positive or negative depending on the experimental conditions, that the pressure-induced transition is more cooperative than the heat-induced transition [Rayan and Macgregor, J Phys Chem B2005, 109, 15558-15565], and that the pressure-induced transition does not occur in the absence of water [Rayan and Macgregor, Biophys Chem, 2009, 144, 62-66]. Additionally, we have shown that ΔV values obtained by pressure-dependent techniques differ from those obtained by ambient pressure techniques such as PPC [Rayan et al. J Phys Chem B2009, 113, 1738-1742] thus shedding light on the effects of pressure on DNA polymers. Herein, we examine the effect of sequence complexity, and hence cooperativity on pressure destabilisation of DNA polymers. Working with Clostridium perfringes DNA under conditions such that the estimated ΔV of the helix-coil transition corresponds to -1.78 mL/mol (base pair) at atmospheric pressure, we do not observe the pressure-induced helix-coil transition of this DNA polymer, whereas synthetic copolymers poly[d(A-T)] and poly[d(I-C)] undergo cooperative pressure-induced transitions at similar ΔV values. We hypothesise that the reason for the lack of pressure-induced helix-coil transition of C. perfringens DNA under these experimental conditions lies in its sequence complexity.

Prospective comparison of breast pain in patients participating in a randomized trial of breast-conserving surgery and tamoxifen with or without radiotherapy.

PURPOSE: To determine whether breast pain affects quality of life (QOL) after breast-conserving surgery and tamoxifen (TAM) with or without adjuvant breast radiotherapy (RT). METHODS AND MATERIALS: A randomized clinical trial was carried out at the Princess Margaret Hospital between 1992 and 2000 to evaluate the need for breast RT in addition to TAM in women >or=50 years treated with breast-conserving surgery for T1-T2N0 breast cancer. A companion study to assess breast pain was carried out during the last 2 years of the randomized clinical trial. The short-form McGill Pain Questionnaire (SF-MPQ), the European Organization for Research and Treatment of Cancer (EORTC) QOL (QLQ-C30) and EORTC breast cancer module (QLQ-BR23) questionnaires were completed by patients within 1 week of randomization in the randomized clinical trial (baseline) and at 3, 6, and 12 months. RESULTS: Eighty-six patients participated in the breast pain study; 41 received RT plus TAM and 45 received TAM alone. The median age was 70 years (range 51-80). The baseline pain and QOL scores were similar for the two groups. No significant difference was found between the two groups for each scale of the QLQ-C30 and QLQ-BR23 questionnaires at 3, 6, or 12 months (p >0.100), except that at 12 months, the score for role function (QLQ-C30) was higher in the RT plus TAM group than in the RT-only group (p = 0.02). At 3 months, the difference between the mean scores for the SF-MPQ was 0.553 (p = 0.47). At 12 months, the pain scores had decreased in both groups; the difference was 0.199 (p = 0.71). The number of breast operations or surgical complications did not correlate with breast pain in either group. Acute RT toxicity scores did not correlate with breast pain or QOL scores at 12 months. CONCLUSION: These results suggest that breast RT does not significantly contribute to breast pain or adversely impact the QOL up to 12 months after treatment in postmenopausal patients with node-negative breast cancer who take TAM.

Pressure-induced helix-coil transition of DNA copolymers is linked to water activity.

We have investigated the effect of reduced water activity on the pressure-stability of double-stranded DNA polymers, poly[d(A-T)] and poly[d(I-C)]. Water activity was modulated by the addition of ethylene glycol and glycerol. The ionic strength of the medium was such that pressure had a destabilising effect on the polymers in the absence of cosolvents. The molar volume change of the heat-induced helix to coil transition (DeltaV(T)) becomes more positive as the activity of water was reduced, suggesting that the pressure-induced denaturation of DNA polymers would not occur at very low water activity. This would imply that water plays a crucial role in the pressure denaturation of DNA, much like that in pressure denaturation of proteins where the driving force of the process is the penetration of water molecules into the protein core [Hummer et al., Proc Natl Acad Sci USA 1998, 95, 1552-1555].