Dispersed lipid liquid crystalline phases stabilized by a hydrophobically modified cellulose

Aqueous dispersions of monoolein (MO) with a commercial hydrophobically modified ethyl hydroxyethyl cellulose ether (HMEHEC) have been investigated with respect to the morphologies of the liquid crystalline nanoparticles. Only very low proportions of HMEHEC are accepted in the cubic and lamellar phases of the monoolein-water system. Due to the broad variation of composition and size of the commercial polymer, no other single-phase regions were found in the quasi-ternary system. Interactions of MO with different fractions of the HMEHEC sample induced the formation of lamellar and reversed hexagonal phases, identified from SAXD, polarization microscopy, and cryogenic TEM examinations. In excess water (more than 90 wt %) coarse dispersions are formed more or less spontaneously, containing particles of cubic phase from a size visible by the naked eye to small particles observed by cryoTEM. At high polymer/MO ratios, vesicles were frequently observed, often oligo-lamellar with inter-lamellar connections. After homogenization of the coarse dispersions in a microfluidizer, the large particles disappeared, apparently replaced by smaller cubic particles, often with vesicular attachments on the surfaces, and by vesicles or vesicular particles with a disordered interior. At the largest polymer contents no proper cubic particles were found directly after homogenization but mainly single-walled defected vesicles with a peculiar edgy appearance. During storage for 2 weeks, the dispersed particles changed toward more well-shaped cubic particles, even in dispersions with the highest polymer contents. In some of the samples with low polymer/MO ratio, dispersed particles of the reversed hexagonal type were found. A few of the homogenized samples were freeze-dried and rehydrated. Particles of essentially the same types, but with a less well-developed cubic character, were found after this treatment. © 2007 American Chemical Society.

Cryopreservation of dog spermatozoa: Effect of bovine serum albumin on acrosomal integrity and pregnancy rates after artificial insemination

The objective of the present study was to compare the in vitro and in vivo profile of frozen dog semen with Tris-bovine serum albumin (TB) and Tris-egg yolk (TE) extenders. Twenty dogs were used as donors. Each dog was stimulated by penile massage and only the sperm-rich fraction was collected weekly until 40 ejaculates were obtained. After macroscopic and microscopic analysis, equal parts of each ejaculate were diluted with TB and TE by the one-step method at 37 °C. The semen was added to 0.5-mL French straws which presented normal characteristics before freezing and after thawing. Acrosomal integrity was evaluated by double Trypan blue-Giemsa staining, in which alive intact (LI), alive reacted (LR), dead intact (DI) and dead reacted (DR) spermatozoa, were identified by the time of thawing and up to 4 h of incubation at 39 °C, the TE being significantly superior to TB (P<0,01) in the LI and LR variables. The TB being significantly superior to TE (P<0,01) in the DR variable. Female dogs in natural heat were submitted to artificial insemination, 20 receiving TE-semen and 20 receiving TB-semen with the Osiris probe (IMV, L'Aigle, France) and the numbers indicate that TE was significantly better than TB (P<0,01) to pregnancy rate and number of puppies/delivery. We concluded from this study, that TE was better than TB, because this, induced an eady acrossome reaction in dog's sperm.

Efeito de um quelante de cálcio, um detergente e da lecitina de soja sobre a qualidade do sêmen caprino congelado-descongelado

The aim of this paper was verifying the effect of the Equex STM Paste and EDTA addition to a Tris-egg yolk extender, on the postthaw goat sperm viability. Nine semen samples of two adult goats were collected by artificial vagina and cryopreserved. It was also objective of this study, to evaluate the utilization of a soybean lecithin based commercial extender (Bioexcell® - IMV, L'Aigle, French) for the goat semen freezing. They were formed five experimental groups: TRIS; TRIS+EDTA; TRIS+EQUEX; TRIS+EDTA+EQUEX e Bioexcell. After evaluation, the semen was diluted in the five extenders and packed in 0.25mL straws with 100 million of motile spermatozoa. The samples were cooled at 0,46°C/min to 5°C, submitted at 75min of equilibration time and frozen in liquid nitrogen vapour. The thawing was accomplished in 37°C water bath for 50s. There were no differences (P>0,05) on the means of post-thaw total and progressive sperm motility among the groups TRIS, TRIS+EQUEX and TRIS+EQUEX+EDTA. The Bioexcell group obtained the least (P<0,05) percentage of post-thawing total and progressive sperm motility. After the thermotolerance test, it was observed the greatest (P<0,05) rates of total and progressive sperm motility in the Equex STM groups (TRIS+EQUEX and TRIS+EQUEX+EDTA). Thus, it can be affirmed that the Equex addition promotes better maintenance rates in the pos-thaw sperm viability, when compared with the extenders that did not contain it.

Treatment of gingival recession with collagen membrane and DFDBA: A histometric study in dogs

In a previous study, we evaluated the findings related to the use of resorbable collagen membranes in humans along with DFDBA (demineralized freeze-dried bone allograft). The aim of this subsequent study was to histometrically evaluate in dogs, the healing response of gingival recessions treated with collagen membrane + DFDBA (Guided Tissue Regeneration, GTR) compared to a coronally positioned flap (CPF). Two types of treatment were randomly carried out in a split-mouth study. Group 1 was considered as test (GTR: collagen membrane + DFDBA), whereas Group 2 stood for the control (only CPF). The dogs were given chemical bacterial plaque control with 0.2% chlorhexidine digluconate during a 90-day repair period. Afterwards, the animals were killed to obtain biopsies and histometric evaluation of the process of cementum and bone formation, epithelial migration and gingival level. A statistically significant difference was found between groups with a larger extension of neoformed cementum (GTR = 32.72%; CPF = 18.82%; p = 0.0004), new bone (GTR = 23.20%; CPF = 09.90%; p = 0.0401) and with a smaller area of residual gingival recession in the test group (GTR = 50.69%; CPF = 59.73%; p = 0.0055) compared to the control group. The only item assessed that showed no statistical difference was epithelial proliferation on the root surface, with means of 15.14% for the GTR group and 20.34% for the CPF group (p = 0.0890). Within the limits of this study we concluded that the treatment of gingival recession defects with GTR, associating collagen membrane with DFDBA, showed better outcomes in terms of a larger extension of neoformed cementum and bone, as well as in terms of a smaller proportion of residual recessions.

Efeito do álcool e formol sobre a composição isotópica de δ 13C e de δ 15N em Plagioscion squamosissimus e Hypophthalmus edentatus (Pisces, Osteichthyes)

The present study investigates the δ 13C and δ 15N isotopic composition in frozen samples (control), samples in alcohol and in formaldehyde of Plagioscion squamosissimus and Hypophthalmus edentatus. From each individual we extracted a strip of muscle from the region above the lateral line, in the dorsal fin base, that was divided into three equal parts, each one was submitted to one type of treatment: freeze - control group (-15oC), conservation in alcohol 70% and fixation in formaldehyde 4%. Samples were kept under those treatments for 30 days, washed and submerged in distilled water for 4 hours. Afterwards, they were dried up in air oven at 60oC for 48 hours and macerated until the obtaining of a fine powder. A significant difference was found in isotopic values of carbon and nitrogen, between the control and the samples in alcohol and formaldehyde, except for δ 13C from the H. edentatus samples in formaldehyde. The carbon isotopic values of samples in alcohol were mostly enriched compared to control, whereas the samples in formaldehyde presented depleted values in relation to the control. The nitrogen isotopic values for both samples preserved in alcohol and formaldehyde were enriched when compared to the values of frozen samples, independently of used preservatives. Therefore, the isotopic correction should be accomplished according to the isotope and preservative employed for species of freshwater fish.

Efecto del método de secado en las propiedades termodinámicas de pulpa de mora en polvo

Three different types of maltodextrin encapsulated dehydrated blackberry fruit powders were obtained using vibrofluidized bed drying (VF), spray drying (SD), vacuum drying (VD), and freeze drying (FD). Moisture equilibrium data of blackberry pulp powders with 18% maltodextrin were determined at 20, 30, 40, and 50°C using the static gravimetric method for the water activity range of 0.06-0.90. Experimental equilibrium moisture content data versus water activity were fit to the Guggenheim-Anderson-de Boer (GAB) model. Agreement was found between experimental and calculated values. The isosteric heat of sorption of water was determined using the Clausius-Clapeyron equation from the equilibrium data; isosteric heats of sorption were found to increase with increasing temperature and could be adjusted by an exponential relationship. For freeze dried, vibrofluidized, and vacuum dried pulp powder samples, the isosteric heats of sorption were lower (more negative) than those calculated for spray dried samples. The enthalpy-entropy compensation theory was applied to sorption isotherms and plots of ΔH versus ΔS provided the isokinetic temperatures, indicating an enthalpy-controlled sorption process.

Use of cooled bull semen as a strategy to increase the pregnancy rate in fixed-time artificial insemination programs-case report

Semen cryopreservation is still considered suboptimal due to lower fertility when compared to fresh semen. The reasons for the loss of fertility are various and related to irreversible damage caused to the cells during the freeze-thaw process. An alternative to conventional cryopreservation represents the use of chilled bull semen, preventing the damage associated with freezing, thereby guaranteeing greater sperm viability. The aim of this study was to describe the use of cooled bull semen as a strategy to increase the pregnancy for Fixed-Time Artificial Insemination (FTAI) of Nellore (Bos indicus) cows. One ejaculate of a select Nellore bull obtained by electroejaculation was used; the semen sample was fractioned into two aliquots: one diluted in Botu-Bov® extender containing 6.4% glycerol for cryopreservation (BB-F, frozen group) and one diluted in the same extender, free from cryoprotectants and used for cooling (BB-C, cooled semen group). The samples in the BB-C group were chilled to 5°C using an isothermic box and maintained for 24 h prior to use. A total of 349 lactating Nellore cows (70-90 days after birth) were synchronized by the insertion of a progesterone releasing device (1.0 g) and estradiol benzoate (2.0 mg i.m.) on a random day of the estrous cycle (Day 0); FTAI was performed 44-48 h after the removal of the device. The pregnancy rates were 45.71 and 61.49% (P<0.05), respectively, for the cryopreserved or chilled bovine semen groups. In conclusion, the use of bull semen cooled for 24 h represents an alternative to conventionally cryopreserved semen, as determined by the increase the pregnancy per artificial insemination in bovine herds. © 2012 Science Publication.

Physicochemical properties of cassava starch oxidized by sodium hypochlorite

In this work, cassava starch was modified by treatment with sodium hypochlorite (NaClO) at different concentrations (0.8, 2.0 and 5.0 % of active chlorine) and selected physicochemical properties of the oxidized starches were investigated. The native and modified samples were evaluated considering moisture, carboxyl content, apparent viscosity, susceptibility to syneresis, mid-infrared spectroscopy and crystallinity index. The treatment with NaClO resulted in alterations in carboxyl content of the oxidized starches that increased with increasing concentration of the oxidant. Oxidized starches also showed higher susceptibility to syneresis, as assessed by the release of liquid during freezing and thawing. Apparent viscosity analysis showed decrease in peak viscosity of the oxidized starches. X-ray diffractograms showed that the oxidation influenced the extent of cassava starch relative crystallinity found to lie between 34.4 % (native) and 39.9 % (2.0 % active chlorine). The infrared spectra are sensitive to structural changes on starch macromolecules and presented characteristic peaks as C-O-C of the six carbon glucose ring absorbs at 1,150-1,085 cm -1 and due to axial deformation these bands changed with the crystal structure of the starch samples. © 2012 Association of Food Scientists & Technologists (India).

Congelação de espermatozoides epididimários de gatos utilizando o diluidor Botu-crio® após refrigeração por 24 h em contêiner de transporte de sêmen Botu-tainer®

The possibilities of using the sperm collected from the epididymis have been widely used because the fertilizing capacity sperm preservation and the possibility of using it for wild cats. But in the process of cryopreservation, some studies show a decrease in the quality of the sperm when left under cooling before frozen for some time. This study aimed to assess the quality of the epididymal sperm obtained from domestic cats after cryopreservation using a diluent based on egg yolk and glycerol (Botu-crio®), comparing the morphofunctional characteristics after cooling for 24 hours in a container of semen transport (Botu-tainer®). We use eight cats submitted to elective orchiectomy, aging from eight months, without racial determination, and good nutritional status. These sperm characteristics were: motility, vigor, concentration, membrane integrity and morphology. It has been found, after statistical analysis, that the container of semen was able to maintain sperm viability, even for 24h. We also observed a significant decrease on all parameters after frozen, consequential, probably to thermal stress that occurs in processing. However, the percentage of membrane integrity after thawing shows good employability of the Botu-crio®, which viability is possible to perform in vitro fertilization, requiring higher ratings.

Polymorphisms in FGFBP1 and FGFBP2 genes associated with carcass and meat quality traits in chickens

In the past, the focus of broiler breeding programs on yield and carcass traits improvement led to problems related to meat quality. Awareness of public concern for quality resulted in inclusion of meat quality traits in the evaluation process. Nevertheless, few genes associated with meat quality attributes are known. Previous studies mapped quantitative trait loci for weight at 35 and 42 days in a region of GGA4 flanked by the microsatellite markers, MCW0240 and LEI0063. In this region, there are 2 fibroblast growth factor binding protein (FGFBP) genes that play an important role in embryogenesis, cellulardifferentiation, and proliferation in chickens. The objective of this study was to identify and associate single nucleotide polymorphisms (SNPs) in FGFBP1 and FGFBP2 with performance, carcass, and meat quality in experimental and commercial chicken populations. In the commercial population, SNP g.2014G>A in FGFBP1 was associated with decreased carcass weight (P < 0.05), and SNP g.651G>A in FGFBP2 was associated with thawing loss and meat redness content (P < 0.05). Four haplotypes were constructed based on 2 SNPs and were associated with breast weight, thawing loss, and meat redness content. The diplotypes were associated with thawing loss, lightness, and redness content. The SNPs evaluated in the present study may be used as markers in poultry breeding programs to aid in improving growth and meat quality traits. © FUNPEC-RP.

Root coverage stability of the subepithelial connective tissue graft and guided tissue regeneration: A 30-month follow-up clinical trial

Objectives: The aim of this study was to compare the long-term clinical effects produced by subepithelial connective tissue graft (SCTG) and guided tissue regeneration combined with demineralized freeze-dried bone allograft (GTR-DFDBA) in the treatment of gingival recessions in a 30-month follow-up clinical trial. Methods: Twenty-four defects were treated in 12 patients who presented canine or pre-molar Miller class I and/or II bilateral gingival recessions. GTR-DFDBA and SCTG treatments were performed in a randomized selection in a split-mouth design. The clinical measurements included root coverage (RC), gingival recession (GR), probing depth (PD), clinical attachment level (CAL) and keratinized tissue width (KTW). These clinical parameters were evaluated at baseline and after 6, 18 and 30 months post-surgery. Results: The changes in RC, GR, PD and CAL did not show significant differences between groups (p > 0.05). Both procedures promoted similar RC (GTR-DFDBA: 87% and SCTG: 95.5%) and similar reduction in GR (GTR-DFDBA: 3.25 mm and SCTG: 3.9 mm), PD (GTR-DFDBA: 1.6 mm and SCTG: 1.2 mm) and CAL (GTR-DFDBA: 4.9 mm and SCTG: 5.0 mm). The increase in KTW was significantly higher (p = 0.02) in the SCTG group (3.5 mm) than in the GTR-DFDBA group (2.4 mm). Conclusions: Both techniques for treatment of gingival recession (SCTG and GTR-DFDBA) lead to favourable and long-term stable results, but SCTG promoted a more favourable increase in keratinized tissue. © 2012 Elsevier Ltd. All rights reserved.

Effect of drying technique on some physical properties of cross-linked high amylose/pectin mixtures

Polymers mixtures as well as cross-linking reactions are approaches that have been used successfully to modulate the polymers characteristics in order to improve the control over drug release rate. High amylose and pectin are polysaccharides frequently used to prepare drug delivery systems. Since the drying technique can strongly influence the properties of such systems, the aim of this work was to characterize high amylose/pectin mixtures cross-linked with sodium trimetaphosphate and dried by different techniques-oven and lyophilization. The results showed that samples dried by lyophilization presented reduced particle size, higher porosity and higher swelling ability than the samples dried in oven. Besides, lower thermal stability and different diffraction patterns showed by the former particles should reflect the structural changes as a function of drying technique. © 2013 Informa Healthcare USA, Inc.

Effect of Storage Time and Temperature of Equine Epididymis on the Viability, Motion Parameters, and Freezability of Epididymal Sperm

The recovery of sperm from the epididymal cauda may be the last chance to obtain genetic material when sudden death or serious injuries occur in valuable stallions. However, the lack of technical knowledge regarding the storage and transportation of the epididymis often prevents the preservation of the sperm. Therefore, the aim of this study was to compare sperm parameters of sperm obtained immediately after orchiectomy with sperm recovered from epididymal cauda at different times after storage at 5°C and at room temperature (RT). For that, 48 stallions of different breeds were used. In group 1 (control group), eight stallions were used, and the harvest of the epididymal sperm was performed immediately after orchiectomy. In group 2, 40 stallions were used, which were divided into five groups according to the storage time of the epididymis after orchiectomy (6, 12, 18, 24, or 30 hours), making a total of eight stallions per group. One epididymis of each stallion was stored at 5°C, and the contralateral epididymis was stored at RT, both for the same period. The sperm parameters of total motility, progressive motility, progressive linear velocity, curvilinear velocity, percentage of rapid sperm, and plasma membrane integrity were evaluated in all the groups after sperm recovery, resuspension in a sperm freezing diluent, and thawing. In conclusion, the storage of the testis-epididymis complex at 5°C provided better preservation of epididymal sperm than the storage at RT, and regardless of the temperature, the progressive motility is the sperm parameter that is most sensitive to storage time. © 2013 Elsevier Inc.

Vitrification of immature feline oocytes with a commercial kit for bovine embryo vitrification

The aim of this study was to evaluate the suitability of a commercial kit for bovine embryo vitrification for cryopreserving cat oocytes and to evaluate comparatively the effects of its use with slow freezing procedure on cryotolerance in terms of morphology and oocyte resumption of meiosis. Germinal vesicle stage oocytes isolated from cat ovaries were either vitrified (n=72) using a vitrification kit for bovine embryo or slow frozen (n=69) by exposing oocyte to ethylene glycol solution before being transferred to a programmable embryo freezer. After thawing and warming, oocytes were cultured for 48h and then were examined for meiosis resumption using bisbenzimide fluorescent staining (Hoechst 33342). Fresh immature oocytes (n=92) were used as the control group. The proportion of oocytes recovered in a morphologically normal state after thawing/warming was significantly higher in frozen oocytes (94.5%) than in the vitrified ones (75%, p<0.01). Morphological integrity after culture was similar in vitrified (73.6%) and slow frozen oocytes (76.8%); however, only 37.5% of the morphologically normal oocytes resumed meiosis after vitrification compared to 60.9% of those submitted to slow freezing procedure (p<0.01). Fresh oocytes showed higher morphological integrity (91.3%) and meiosis resumption rates (82.6%, p<0.002) than cryopreserved oocytes, irrespective of the procedure used. These results suggest that immature cat oocytes vitrified with a kit for bovine embryos retain their capacity to resume meiosis after warming and culture, albeit at lower rates than slow frozen oocytes. Vitrification and slow freezing methods show similar proportions of oocytes with normal morphology after culture, which demonstrate that thawed and warmed oocytes that resist to cryodamage have the same chances to maintain their integrity after 48h of culture. © 2012 Blackwell Verlag GmbH.

Proteômica para investigar a congelabilidade do semen bovino

The present study was undertaken the protein composition in 2D-electrophoretic pattern (2DE) of the seminal plasma (SP) can interfere in the semen bull freezability, and if we can use that for predicting semen bull freezability. Samples were obtained of 20 bulls (different breeds) with a minimum of 3 years history semen production in commercial semen collection Center. All animals ranged between 2 - 7 years of age. The semen freezability was calculated by # of thawed and approved ejaculates / # of ejaculates submitted to cryopreservation (after semen evaluation and approved to submitted to freeze). The bulls were divided in 3 groups: HIGH (=>80% ejaculates approved); MEDIUM (>60% and <79% ejaculates approved); LOW (=<59% ejaculates approved); the pattern and criteria were the same used in the routine of the commercial semen Center. 68 gels were carried through by 2DE of SP samples indicated 225 detected spots with protein different amount (VION) comparing. Comparing bull ́s semen freezability and VION of each spot found difference among 2 spots from High and Low, even considering just spots with % of detection frequency bigger than 75%. The taurine bulls demonstrated more homogeneous profile when comparing with zebu bulls, considering number and frequency of appearance of spots. The results showed that proteomics can be a useful tool to predict the semen freezability, but we ́ll need to study better the interactions between sperm membrane, seminal plasma and extender to comprehend better which proteome phenotype interfere positive or negatively in the semen freezability.