Search results for: sperm viability

Authors: Mengfan Li, Filip Van Bockstaele, Wenyong Lou, Frank Devlighere

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The development of probiotics-encapsulated emulsions that maintain the viability of probiotics during processing, storage and human gastrointestinal (GI) tract environment receives great scientific and commercial interest. In this study, typical W/O and O/W emulsions with and without oil gelation were used to encapsulate L. plantarum. The effects of emulsion types on the viability of L. plantarum during storage and GI tract were investigated. Besides, the position of L. plantarum in emulsion system and its number of viable cells when threating by adverse environment was correlated in order to figure out which type of emulsion is more suitable as food carrier for probiotics encapsulation and protection. As a result, probiotics tend to migrate from oil to water phase due to the natural hydrophilicity; however, it’s harmful for cells viability when surrounding by water for a long time. Oil gelation in emulsions is one of the promising strategies for inhibiting the cells mobility and decreasing the contact with adverse factors (e.g., water, exogenous enzymes and gastric acid), thus enhancing the number of viable cells that enough to exert its beneficial effects in host.

Keywords: emulsion, gelation, encapsulation, probiotics

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Authors: A. G. Miah, U. Salma, K. Schellander

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Relaxin was first described in 1926 by Frederick Hisaw. Previously it was considered as only the hormone of pregnant mammals due to its important roles in pregnancy and parturition. From the last decade, the physiological role of relaxin in male reproduction has been given experimental attention, and the results have made it clear that relaxin can no longer be considered strictly as only the hormone of female reproduction. The accessory glands (specially, the prostate glands) of the male reproductive system are the source of seminal relaxin, which is secreted into the seminal plasma and saturated with spermatozoa just after ejaculation. Several studies have reported that relaxin has important roles in improving motility in human sperm. Thereafter, the growing interest on relaxin has intensified efforts to investigate the role of relaxin in other sperm physiological phenomena like, capacitation, acrosome reaction, and their mediating factors associated with successful fertilization. Therefore, this review aims to provide up-to-date information about the physiological roles of relaxin in sperm motility, capacitation, acrosome reaction, and their mediating factors, such as, utilization of glucose, cholesterol efflux, Ca2+-influx, intracellular cAMP and protein tyrosine phosphorylation. Some studies have shown relaxin to increase the percentage of progressive motility and induce capacitation and acrosome reaction through increasing the utilization of glucose and mediating the cholesterol efflux, Ca2+-influx, intracellular cAMP and protein tyrosine phosphorylation. Thus, the review suggests that the supplementation of relaxin into the capacitating medium may contribute the possible beneficial roles in fresh and cryopreserved spermatozoal prefertilization events.

Keywords: relaxin, physiological roles, prefertilizing activities, spermatozoa

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Authors: Gloria Pinilla, Jose Manuel Baena, Patricia Gálvez-Martín, Juan Antonio Marchad

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Cartilage is a dense connective tissue with limited self-repair properties. Currently, the therapeutic use of autologous or allogenic chondrocytes makes up an alternative therapy to the pharmacological treatment. The design of a bioprinted 3D cartilage with chondrocytes and biodegradable biomaterials offers a new therapeutic alternative able of bridging the limitations of current therapies in the field. We have developed an enhanced printing processes-Injection Volume Filling (IVF) to increase the viability and survival of the cells when working with high-temperature thermoplastics without the limitation of the scaffold geometry in contact with cells. We have demonstrated the viability of the printing process using chondrocytes for cartilage regeneration. This development will accelerate the clinical uptake of the technology and overcomes the current limitation when using thermoplastics as scaffolds. An alginate-based hydrogel combined with human chondrocytes (isolated from osteoarthritis patients) was formulated as bioink-A and the polylactic acid as bioink-B. The bioprinting process was carried out with the REGEMAT V1 bioprinter (Regemat 3D, Granada-Spain) through a IVF. The printing capacity of the bioprinting plus the viability and cell proliferation of bioprinted chondrociytes was evaluated after five weeks by confocal microscopy and Alamar Blue Assay (Biorad). Results showed that the IVF process does not decrease the cell viability of the chondrocytes during the printing process as the cells do not have contact with the thermoplastic at elevated temperatures. The viability and cellular proliferation of the bioprinted artificial 3D cartilage increased after 5 weeks. In conclusion, this study demonstrates the potential use of Regemat V1 for 3D bioprinting of cartilage and the viability of bioprinted chondrocytes in the scaffolds for application in regenerative medicine.

Keywords: cartilage regeneration, bioprinting, bioink, scaffold, chondrocyte

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Authors: Mayuva Youngsabanant, Suphaphorn Rabiab, Hatairuk Tungkasen, Nongnuch Gumlungpat, Mayuree Pumipaiboon

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The porcine follicular fluid proteins (pFF) of healthy small size ovarian follicles (1-3 mm in diameters) of Large White pig ovaries were collected by sterile technique. They were used for testing the effect on cell viability and characterization of Vero cell lines using MTT assay. Two hundred microliter of round shape Vero cell lines were culture in 96 well plates with DMEM for 24 h. After that, they were attachment to substrate and some changed into fibroblast shape and spread over the surface after culture for 48 h. Then, Vero cell lines were treated with pFF at concentration of 2, 4, 20, 40, 200, 400, 500, and 600 µg proteins/mL for 24 h. Yields of the best results were analyzed by using one-way ANOVA. MTT assay reviewed an increasing in percentage of viability of Vero cell lines indicated that at concentration of 400-600 µg proteins/mL showed higher percentage of viability (115.64 ± 6.95, 106.91 ± 5.27 and 116.73 ± 20.15) than control group. They were significantly different from the control group (p < 0.05) but lower than the positive control group (DMEM with 10% heat treated fetal bovine serum). Cell lines showed normal character in fibroblast elongate shape after treated with pFF except in high concentration of pFF. This result implies that pFF of small size ovarian follicle at concentration of 400-600 µg proteins/mL could be optimized concentration for using as a supplement in Vero cell line culture medium to promote cell viability instead of growth hormone from fetal bovine serum. This merit could be applied in other cell biotechnology researches. Acknowledgements: This work was funded by a grant from Silpakorn University and Faculty of Science, Silpakorn University, Thailand.

Keywords: cell viability, porcine follicular fluid, MTT assay, Vero cell line

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Authors: Benbellil-Tafoughalt Saida, Tababouchet Meriem

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Sexual selection is the most supported explanation for genital extravagance occurring in animals. In promiscuous species, population density, as well as climate conditions, may act on the sperm competition intensity, one of the most important mechanism of post-copulatory sexual selection. The present study is empirical testing of sexual selection's potential role on genitalia variation in the simultanuously hermaphroditic land snail Helixaperta (Pulmonata, Stylommatophora). The purpose was to detect the patterns as well as the origin of the distal genitalia variability and especially to test the potential effect of sexual selection. The study was performed on four populations, H. aperta, different in habitat humidity regimes and presenting variable densities, which were mostly low. The organs of interest were those involved in spermatophore production, reception, and manipulation. We examined whether the evolution of those organs is connected to sperm competition intensity which is traduced by both population density and microclimate humidity. We also tested the hypothesis that those organs evolve in response to shell size. The results revealed remarkable differences in both snails’ size and organs lengths between populations. In most cases, the length of genitalia correlated positively to snails’ body size. Interestingly, snails from the more humid microclimate presented the highest mean weight and shell dimensions comparing to those from the less humid microclimate. However, we failed to establish any relation between snail densities and any of the measured genitalia traits.

Keywords: fertilization pouch, helix aperta, land snails, reproduction, sperm storage, spermatheca

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Authors: M. M. Islam, S. Sharmin, M. Shah Newaz, N. S. Juyena, M. M. Rahman, P. K. Jha, F. Y. Bari

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The aim of the present study was to select the high quality ram by measuring the scrotal biometry which has an effect on semen parameters. Ten rams were selected in the present study. Eight ejaculates were collected from each ram using artificial vagina method. Scrotal circumference was measured before and after semen collection on weekly basis using the Scrotal tape. Bio-metries of scrotum (scrotal length and scrotal volume) were calculated. Semen was evaluated for macroscopic and microscopic characteristics. The average estimated scrotal circumference (cm) and scrotal volume (cm3) in 8 different age groups were 17.16±0.05 cm and 61.30±0.70 cm3, 17.17±0.62 cm and 63.67±4.49 cm3, 17.22±0.52 cm and 64.90±4.21 cm3, 17.72±0.37 cm and 67.10±4.20 cm3, 18.41±0.35cm and 69.52±4.12cm3, 18.45±0.36cm and 77.17±3.81 cm3, 18.55±0.41 cm and 78.72±4.90 cm3, 19.10±0.30 cm and 87.35±5.45 cm3 respectively. The body weight, scrotal circumference and scrotal volume increased with the progress of age (P < 0.05). Body weight of age group 381-410 days (13.62+1.48 kg) was significantly higher than group 169-200 days (10.17±0.05 kg) and 201-230 days (10.42±1.18 kg) (p < 0.05). Scrotal circumference (SC) of age group 381-410 days (19.10±0.30 cm) was significantly higher (p < 0.05) than other groups. In age group 381-410 days, scrotal volume (SCV) (87.35±5.45 cm3) was significantly higher than other first five groups (p < 0.05). Both scrotal circumference and scrotal volume development was positively correlated with the increasing of body weight (R2= 0.51). Semen volume increased accordingly with the increasing of ages, varied from 0.35±0.00 ml to 1.15+0.26 ml. Semen volume of age group 381-410 days (1.15±0.26 ml) was significantly higher than other age groups (p < 0.05) except age group 351-380 days (p > 0.05). Mass activity of different age groups varied from 2.75 (±0.35) to 4.25 (±0.29) ml in the scale of 1-5. Sperm concentration, progressive motility (%),progressively improved according to the increasing of ages, but significant changes in these parameters were seen when the animals reaches the age 291 days or more (p < 0.05). However, normal spermatozoa (%) improved significantly from the age of 261 days or more. Mass activity (mass) was positively correlated with sperm concentration (R2=0.568) and progressive motility (%) (R2=0.616). The relationships of semen volume with body weight and scrotal measurements and sperm concentration indicate that they are useful in evaluating rams for breeding soundness and genetic improvement for fertility in indigenous ram.

Keywords: breeding soundness, ram, semen quality, scrotal biometry

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Authors: Kamal Kumar Gupta

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The present research work was carried out on Dysdercus koenigii to understand various aspects of reproductive behavior such as mate finding and recognition, mate selection and mating preference, mating receptivity, and prolonged copulation. The studies carried out on mate searching and courtship behaviour of Dysdercus reflected the courtship behaviour in Dysdercus was brief. The opposite sexes are brought together by the pheromone. The males responded to female sex pheromones by showing directional movements toward the sex partners. Change in mating receptivity pattern of female Dysdercus was ascertained using three parameters of mating behaviour i.e. numbers of male’s encounter, the time taken to mate successfully and per cent females responding to mating. It was seen that a receptive female responded positively to the courting males and a high percentage of females mate usually in a very short time span. The females of Dysdercus showed continued mating receptivity throughout their life. The studies pertaining to mate selection by females showed that females generally do not discriminate among males and usually mate with any male they encountered first. The adults of Dysdercus remain in continuous copula up to 72hr. and mate 5-7 time in their life span. Studies pertaining to significance of prolonged mating in the life time reproductive success of the female Dysdercus indicated that fecundity and fertility and oviposition behavior of the female Dysdercus was related to duration of mating. In order to understand sperm precedence, the sterilized males were produced by exposing them to Gamma radiation. Our studies indicated that a dose of 50 Gy of Gamma radiations induced 95% sterility but does not impair the mating behaviour drastically. To understand role of sperms which were transfer during second mating in fertilizing the subsequent egg batches the sperm utilization pattern of doubly mated female was assessed. The females were mated with normal male or sterilized male in a combination. The sperm utilization pattern was determined by P2 value, our studies indicated a very high P2 value of 0.966, and indicated that sperms of last mating were utilized by the female for fertilization. In light of some of the unique reproductive behaviour of Dysdercus koenigii, such as brief courtship behavior, generalized mate selection by the female, continued mating receptivity and a prolonged pre oviposition period, the present studies on sperm precedence provides an explanation to an unusually prolonged copulation in Dysdercus.

Keywords: dysdercus koenigii, mating behaviour, reproductive performance, entomology

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Authors: Eva Tvrdá, Hana Greifová, Peter Ivanič, Norbert Lukáč

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The aim of this study was to evaluate the dose- and time-dependent in vitro effects of berberine (BER), a natural alkaloid with numerous biological properties on bovine spermatozoa during three time periods (0 h, 2 h, 24 h). Bovine semen samples were diluted and cultivated in physiological saline solution containing 0.5% DMSO together with 200, 100, 50, 10, 5, and 1 μmol/L BER. Spermatozoa motility was assessed using the computer assisted semen analyzer. The viability of spermatozoa was assessed by the metabolic (MTT) assay, production of superoxide radicals was quantified using the nitroblue tetrazolium (NBT) test, and chemiluminescence was used to evaluate the generation of reactive oxygen species (ROS). Cell lysates were prepared and the extent of lipid peroxidation (LPO) was evaluated using the TBARS assay. The results of the movement activity showed a significant increase in the motility during long term cultivation in case of concentrations ranging between 1 and 10 μmol/L BER (P < 0.01; P < 0.001; 24 h). At the same time, supplementation of 1, 5 and 10 μmol/L BER led to a significant preservation of the cell viability (P < 0.001; 24 h). BER addition at a range of 1-50 μmol/L also provided a significantly higher protection against superoxide (P < 0.05) and ROS (P < 0.001; P < 0.01) overgeneration as well as LPO (P < 0.01; P<0.05) after a 24 h cultivation. We may suggest that supplementation of BER to bovine spermatozoa, particularly at concentrations ranging between 1 and 50 μmol/L, may offer protection to the motility, viability and oxidative status of the spermatozoa, particularly notable at 24 h.

Keywords: berberine, bulls, motility, oxidative profile, spermatozoa, viability

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Authors: Stephen Adeniyi Adefegha, Vitor Mostardeiro, Vera Maria Morsch, Ademir F. Morel, Ivana Beatrice Manica Da Cruz, Sabrina Somacal Maria Rosa Chitolina Schetinger

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Moringa stenopetala is often consumed as food and used in folkloric medicine for the management of several diseases. Purpose: This study was set up in order to assess the effect of aqueous extract of Moringa stenopetala on cell viability and oxidative stress biomarkers in BV-2 microglial cells. Aqueous extracts of M. stenopetala were prepared, lyophilized and reconstituted in 0.5% dimethylsulphoxide (DMSO). Cells were treated with M. stenopetala extracts (0.1 - 100 µg/ml) for cell viability and nitric oxide (NO) production tests. However, M. stenopetala extract (50 µg/ml) was used in the treatment of cells for the determination of protein carbonyl content and reactive oxygen species (ROS) level. Incubation of BV-2 microglia cell with M. stenopetala extract maintained cell viability, diminished NO and ROS levels, and reduced protein carbonyl contents Chlorogenic acid, rutin, kaempferol and quercetin derivatives were the main phenolic compounds identified in M. stenopetala leaf extract. These phenolic compounds present in M. stenopetala may be responsible for the mitigation of oxidative stress in BV-2 microglial cells.

Keywords: oxidative stress, BV-2 microglial cell, Moringa stenopetala, cell viability, antioxidant

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Authors: Geetika Richa, M. L. Sharma

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Bamboo as an ancient herbal medicine has been used for thousands of years in Asia and goes by many names such as tabashir, banslochan etc. It is often used for its tonic and astringent properties. Modern analysis of bamboos show high amount of vitamins and minerals which makes them valuable as a curative. Bamboo leaf decoction and young shoots are known as remedy for intestinal worms, healing of ulcers and stomach disorders. Bamboos are known to be propagated by large scale plantations but propagation through seeds occurs very limited as they have very short viability of few months. Seeds loses viability over a period of time even under controlled conditions and important factors that affect seed viability is the decline in reserve food material, decrease in membrane integrity and fall in endogenous level of growth hormones. Invigoration treatments that include hydration, dehydration, incorporation of bioactive chemicals such as growth regulators, nutrients and antioxidants etc. improve the seed performance. Our studies were aimed to determine the most effective invigoration treatments to enhance vigour and viability of seeds by following invigoration treatments, i.e., hardening. Treated seeds were stored at controlled temperature and humidity (in desiccators at 4°C). In hardening, chemicals were applied in 3 different concentrations to three replicates of 10 seeds. Hardening was done withGA3, IAA, (each with concentrations of 10 ppm, 20 ppm and 50 ppm), calcium oxychloride, neem leaf powder and clay (each with concentrations of 2%, 5% and 10%). Statistically all the hardening materials were effective but GA3 50 ppm was the most effective one in maintaining germination percentage and vigour index. Hardening treatments increased the germination percentage of seeds, i.e. 86.2%, over control which showed germination percentage of 80.2%. It was concluded that in order to maintain seed viability during storage for longer period of time, invigoration treatments have been found to be very effective.

Keywords: invigoration, seed quality, viability, hardening, membrane integrity, decoction

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Authors: L. A. Bosnea, T. Moschakis, C. Biliaderis

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Two probiotic strains of Lactobacillus paracasei subsp. paracasei (E6) and Lactobacillus paraplantarum (B1), isolated from traditional Greek dairy products, were microencapsulated by complex coacervation using whey protein isolate (WPI, 3% w/v) and gum arabic (GA, 3% w/v) solutions mixed at different polymer ratio (1:1, 2:1 and 4:1). The effect of total biopolymer concentration on cell viability was assessed using WPI and GA solutions of 1, 3 and 6% w/v at a constant ratio of 2:1. Also, several parameters were examined for optimization of the microcapsule formation, such as inoculum concentration and the effect of ionic strength. The viability of the bacterial cells during heat treatment and under simulated gut conditions was also evaluated. Among the different WPI/GA weight ratios tested (1:1, 2:1, and 4:1), the highest survival rate was observed for the coacervate structures made with the ratio of 2:1. The protection efficiency at low pH values is influenced by both concentration and the ratio of the added biopolymers. Moreover, the inoculum concentration seems to affect the efficiency of microcapsules to entrap the bacterial cells since an optimum level was noted at less than 8 log cfu/ml. Generally, entrapment of lactobacilli in the complex coacervate structure enhanced the viability of the microorganisms when exposed to a low pH environment (pH 2.0). Both encapsulated strains retained high viability in simulated gastric juice (>73%), especially in comparison with non-encapsulated (free) cells (<19%). The encapsulated lactobacilli also exhibited enhanced viability after 10–30 min of heat treatment (65oC) as well as at different NaCl concentrations (pH 4.0). Overall, the results of this study suggest that complex coacervation with WPI/GA has a potential to deliver live probiotics in low pH food systems and fermented dairy products; the complexes can dissolve at pH 7.0 (gut environment), releasing the microbial cells.

Keywords: probiotic, complex coacervation, whey, encapsulation

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Authors: Farideh Tabatabei-Yazdi, Fereshteh Falah, Alireza Vasiee

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Nowadays, production of functional foods has become very essential. Inulin is one of the most functional hydrocolloid compounds used in such products. In the present study, the production of a synbiotic yogurt containing 1, 2.5, and 5% (w/v) inulin has been investigated. The yogurt was fermented with Lactobacillus brevis PML1 derived from Tarkhineh, an Iranian cereal-dairy fermented food. Furthermore, the physicochemical properties, antioxidant activity, sensory attributes, and microbial viability properties were investigated on the 0th, 7th, and 14th days of storage after fermentation. The viable cells of L. brevis PML1 reached 108 CFU/g, and the product resisted to simulated digestive juices. Moreover, the synbiotic yogurt impressively increased the production of antimicrobial compounds and had the most profound antimicrobial effect on S. typhimurium. The physiochemical properties were in the normal range, and the fat content of the synbiotic yogurt was reduced remarkably. The antioxidant capacity of the fermented yogurt was significantly increased (p<0:05), which was equal to those of DPPH (69:18±1:00%) and BHA (89:16±2:00%). The viability of L. brevis PML1 was increased during storage. Sensory analysis showed that there were significant differences in terms of the impressive parameters between the samples and the control (p<0:05). Addition of 2.5% inulin not only improved the physical properties but also retained the viability of the probiotic after 14 days of storage, in addition to the viability of L. brevis with a viability count above 6 log CFU/g in the yogurt. Therefore, a novel synbiotic product containing L. brevis PML1, which can exert the desired properties, can be used as a suitable carrier for the delivery of the probiotic strain, exerting its beneficial health effects.

Keywords: functional food, lactobacillus brevis, symbiotic yogurt, physiochemical properties

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Authors: I. Wiedemann, A. R. Sharifi, A. Mählmeyer, C. Knorr

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A requirement for sperm motility is a morphologically intact flagellum with a central axoneme. The flagellar beating is caused by the varying activation and inactivation of dynein molecules which are located in the axoneme. DNAL4 (dynein, axonemal, light chain 4) is regarded as a possible functional candidate gene encoding a small subunit of the dyneins. In the present study, 5814bp of the porcine DNAL4 (GenBank Acc. No. AM284696.1, 6097 bp, 4 exons) were comparatively sequenced using three boars with a high motility (>68%) and three with a low motility (<60%). Primers were self-designed except for those covering exons 1, 2 and 3. Prior to sequencing, the PCR products were purified. Sequencing was performed with an ABI PRISM 3100 Genetic Analyzer using the BigDyeTM Terminator v3.1 Cycle Sequencing Reaction Kit. Finally, 23 SNPs were described and genotyped for 82 AI boars representing the breeds Piétrain, German Large White and German Landrace. The genotypes were used to assess possible associations with standard spermatological parameters (ejaculate volume, density, and sperm motility (undiluted (Motud), 24h (Mot1) and 48h (Mot2) after semen collection) that were regularly recorded on the AI station. The analysis included a total of 8,833 spermatological data sets which ranged from 2 to 295 sets per boar in five years. Only SNP g.1007A>G had a significant effect. Finally, the gene substitution effect using the following statistical model was calculated: Yijk= µ+αi+βj+αβij+b1Sijk+b2Aijk+b3T ijk + b4Vijk+b5(α*A)ijk +b6(β*A)ijk+b7(A*T)ijk+Uijk+eijk where Yijk is the semen characteristics, µ is the general mean, α is the main effect of breed, β is the main effect of season, S is the effect of SNP (g.1007A > G), A is the effect of age at semen collection, V is the effect of diluter, αβ, α*A, β*A, A*T are interactions between the fixed effects, b1-b7 are regression coefficients between y and the respective covariate, U is the random effect of repeated observation on animal and e is the random error. The results from the single marker regression analysis revealed highly significant effects (p < 0.0001) of SNP g.1007A > G on Mot1 resp. on Mot2, resulting in a marked reduction by 11.4% resp. 15.4%. Furthermore a loss of Motud by 4.6% was detected (p < 0.0178). Considering the SNP g.1007A > G as a main factor (dominant-recessive model), significant differences between genotypes AA and AG as well as AA and GG for Mot1 and Mot2 exist. For Motud there was a significant difference between AA and GG.

Keywords: association, DNAL4, porcine, sperm traits

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Authors: Sonal Gupta, Rakhi Bansal, Javed Ali, Reema Gabrani, Shweta Dang

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Nanoemulsions (NEs) and microemulsions (MEs) have been an attractive tool for encapsulation of both hydrophilic and lipophillic actives. Both these systems are composed of oil phase, surfactant, co-surfactant and aqueous phase. Depending upon the application and intended use, both oil-in-water and water-in-oil emulsions can be designed. NEs are fabricated using high energy methods employing less percentage of surfactant as compared to MEs which are self assembled drug delivery systems. Owing to the nanometric size of the droplets these systems have been widely used to enhance solubility and bioavailability of natural as well as synthetic molecules. The aim of the present study is to assess the effect of % age of surfactants on cell viability of Vero cells (African Green Monkeys’ Kidney epithelial cells) via MTT assay. Green tea catechin (Polyphenon 60) loaded ME employing low energy vortexing and NE employing high energy ultrasonication were prepared using same excipients (labrasol as oil, cremophor EL as surfactant and glycerol as co-surfactant) however, the % age of oil and surfactant needed to prepare the ME was higher as compared to NE. These formulations along with their excipients (oilME=13.3%, SmixME=26.67%; oilNE=10%, SmixNE=13.52%) were added to Vero cells for 24 hrs. The tetrazolium dye, 3-(4,5-dimethylthia/ol-2-yl)-2,5-diphi-iiyltclrazolium bromide (MTT), is reduced by live cells and this reaction is used as the end point to evaluate the cytoxicity level of a test formulation. Results of MTT assay indicated that oil at different percentages exhibited almost equal cell viability (oilME ≅ oilNE) while surfactant mixture had a significant difference in the cell viability values (SmixME < SmixNE). Polyphenon 60 loaded ME and its PlaceboME showed higher toxicity as compared to Polyphenon 60 loaded NE and its PlaceboNE that can be attributed to the higher concentration of surfactants present in MEs. Another probable reason for high % cell viability of Polyphenon 60 loaded NE might be due to the effective release of Polyphenon 60 from NE formulation that helps in the sustenance of Vero cells.

Keywords: cell viability, microemulsion, MTT, nanoemulsion, surfactants, ultrasonication

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Authors: Wendy Kramer

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How and when a donor-conceived person (DCP) learns about their conception significantly affects their experiences and choices, including whether they'd consider using a donor or donating their own gametes. Objective: We sought to identify factors that positively and negatively impact the experience of being a DCP. We sought to determine if DCP would consider utilizing donor gametes themselves, if unable to conceive spontaneously and if DCP were likely to be donors themselves. Materials and Methods: A cross-sectional survey of adult DCP was disseminated to members of the Donor Sibling Registry. The survey consisted of 31 items including whether experience as DCP was positive or negative, the willingness to use donor gametes if spontaneous conception was not an option, and questions regarding donating gametes. Results: 529 people (81.7% female) completed the survey, the median age was 28 years (range 18-77 years) and 94.7% were conceived via donor sperm. Most felt "neutral" (31.6%), "positive" (26.3%) or "very positive" (20.8%) about being a DCP regardless of donor type. While most found out about being a DCP after age 18 (63.4%), those with a positive experience were more likely to "have always known" (40.7%). Conclusions: People conceived by donor-assisted reproduction are more likely to have neutral to positive overall feelings surrounding their conception if they are told at a very young age about their donor-conceived origins by a family member. The majority of DCP are willing to adopt but would not consider using donated gametes themselves if unable to conceive spontaneously. DCP are not likely to become donors themselves despite the majority of DCP having a high positive feeling regarding being donor-conceived.

Keywords: donor conception, donor offspring, sperm donation, egg donation, donor-conceived people

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Authors: Yuxiang Pan, Yong Qiu, Chenlei Gu, Ping Wang

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In the past decade, three-dimensional (3D) tumor cell models have attracted increasing interest in the field of drug screening due to their great advantages in simulating more accurately the heterogeneous tumor behavior in vivo. Drug sensitivity testing based on 3D tumor cell models can provide more reliable in vivo efficacy prediction. The gold standard fluorescence staining is hard to achieve the real-time and label-free monitoring of the viability of 3D tumor cell models. In this study, micro-groove impedance sensor (MGIS) was specially developed for dynamic and non-invasive monitoring of 3D cell viability. 3D tumor cells were trapped in the micro-grooves with opposite gold electrodes for the in-situ impedance measurement. The change of live cell number would cause inversely proportional change to the impedance magnitude of the entire cell/matrigel to construct and reflect the proliferation and apoptosis of 3D cells. It was confirmed that 3D cell viability detected by the MGIS platform is highly consistent with the standard live/dead staining. Furthermore, the accuracy of MGIS platform was demonstrated quantitatively using 3D lung cancer model and sophisticated drug sensitivity testing. In addition, the parameters of micro-groove impedance chip processing and measurement experiments were optimized in details. The results demonstrated that the MGIS and 3D cell-based biosensor and would be a promising platform to improve the efficiency and accuracy of cell-based anti-cancer drug screening in vitro.

Keywords: micro-groove impedance sensor, 3D cell-based biosensors, 3D cell viability, micro-electromechanical systems

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Authors: Vikas Sharma, V. K. Dixit

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Anacyclus pyrethrum (A. pyrethrum) has been used as Vajikaran Rasayana (aphrodisiac) in traditional Indian ayurvedic medicine to treat male sexual dysfunction, including infertility. Aphrodisiac activity may be due to an increase in the production or effect of androgens, so this study sought to evaluate the androgenic and spermatogenic potential of the alkylamide-rich ethanol solution extract. Male Wistar strain rats weighing between 150 and 180 g were completely randomized divided into five groups. The ethanol solution extract of A. pyrethrum was administered to groups of rats in 50, 100, and 150 mg/kg doses for a period of 28 days, and the action was compared with control and testosterone-treated rats. Thirteen N-alkylamides were detected in the extract by using HPLC/UV/electrospray ionization mass spectrometry method. Extract administration at all the doses produced significant increase in body weight, sperm count, motility, and viability along with serum testosterone, luteinizing hormone, and follicle-stimulating hormone concentrations. Histoarchitecture of testis revealed increased spermatogenic activities. Seminal fructose content was also significantly increased after 28 days of treatment. Our results suggest that the ethanol solution extract of the roots of A. pyrethrum has androgenic potential and may improve male fertility by enhancing spermatogenesis.

Keywords: N-alkylamides, testosterone, Anacyclus pyrethrum, androgen

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Authors: Dimitrios J. Dimitriou, Maria F. Sartzetaki

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Transport infrastructures are high-cost, long-term investments that serve as vital foundations for the operation of a region or nation and are essential to a country’s or business’s economic development and prosperity, by improving well-being and generating jobs and income. The development of appropriate financing options is of key importance in the decision making process in order develop viable transport infrastructures. The development of transport infrastructure has increasingly been shifting toward alternative methods of project financing such as Public Private Partnership (PPPs) and hybrid forms. In this paper, a methodological decision-making framework based on the evaluation of the financial viability of transportation infrastructure for different financial schemes is presented. The framework leads to an assessment of the financial viability which can be achieved by performing various financing scenarios analyses. To illustrate the application of the proposed methodology, a case study of rail transport infrastructure financing scenario analysis in Greece is developed.

Keywords: rail transport infrastructure, financial viability, scenario analysis, rail project feasibility

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Authors: Zohreh Didar Yaser, Zanganeh Asadabadi

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The aim of this work is to open up the structure of lemon peel (Citrus × latifolia) with mild pretreatments. The effects of autoclave, microwave and ultrasonic with or without acid addition were investigated on the amount of glucose, soluble and insoluble lignin, furfural, yeast viability and bioethanol. The finding showed that autoclave- acid impregnated sample, has the highest glucose release from lignocellulose materials (14.61 and 14.95 g/l for solvent exposed and untreated sample, respectively) whereas at control sample glucose content was at its minimal level. Pretreatments cause decrease on soluble and insoluble lignin and the highest decrease cause by autoclave following with microwave and ultrasonic pretreatments (p≤5%). Moderate increase on furfural was seen at pretreated samples than control ones. Also, the most yeast viability and bioethanol content was belong to autoclave samples especially acid- impregnated ones (40.33%). Comparison between solvent treated and untreated samples indicated that significant difference was between two tested groups (p≤1%) in terms of lignin, furfural, cell viability and ethanol content but glucose didn’t show significant difference. It imply that solvent extraction don’t influences on glucose release from lignocellulose material of lemon peel but cause enhancement of yeast viability and bioethanol production.

Keywords: Bioethanol, Lemon peel, Pretreatments, Solvent Extraction

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Authors: Wendy Kramer

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How and when a donor-conceived person (DCP) learns about their conception significantly affects their experiences and choices, including whether they'd consider using a donor or donating their own gametes. Objective: We sought to identify factors that positively and negatively impact the experience of being a DCP. We sought to determine if DCP would consider utilizing donor gametes themselves, if unable to conceive spontaneously and if DCP were likely to be donors themselves. Materials and Methods: A cross-sectional survey of adult DCP was disseminated to members of the Donor Sibling Registry. The survey consisted of 31 items, including whether experience as DCP was positive or negative, the willingness to use donor gametes if spontaneous conception was not an option, and questions regarding donating gametes. Results: 529 people (81.7% female) completed the survey, the median age was 28 years (range 18-77 years), and 94.7% were conceived via donor sperm. Most felt "neutral" (31.6%), "positive" (26.3%) or "very positive" (20.8%) about being a DCP regardless of donor type. While most found out about being a DCP after age 18 (63.4%), those with a positive experience were more likely to "have always known" (40.7%). Conclusions: People conceived by donor-assisted reproduction are more likely to have neutral to overall positive feelings surrounding their conception if they are told at a very young age about their donor-conceived origins by a family member. The majority of DCPs are willing to adopt but would not consider using donated gametes themselves if unable to conceive spontaneously. DCPs are not likely to become donors themselves despite the majority of DCP having a high positive feeling regarding being donor-conceived.

Keywords: donor conception, sperm donation, oocyte donation, donor-conceived people, infertility

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Authors: Leda Maria Costa Pereira, Maria Denise Lopes, Nucharin Songsasen

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Mammalian ovaries contain thousands of follicles that eventually degenerate or die after culture in vitro. Caspase-3 is a key enzyme that regulating cell death. Our objective was to examine the influence of anti-apoptotic drug Z-VAD-FMK (pan-caspase inhibitor) on in vitro viability of dog follicles within the ovarian cortex. Ovaries were obtained from prepubertal (age, 2.5–6 months) and adult (age, 8 months to 2 years) bitches and ovarian cortical fragments were recovered. The cortices were then incubated on 1.5% (w/v) agarose gel blocks within a 24-wells culture plate (three cortical pieces/well) containing Minimum Essential Medium Eagle - Alpha Modification (Alpha MEM) supplemented with 4.2 µg/ml insulin, 3.8 µg/ml transferrin, 5 ng/ml selenium, 2 mM L-glutamine, 100 µg/mL of penicillin G sodium, 100 µg/mL of streptomycin sulfate, 0.05 mM ascorbic acid, 10 ng/mL of FSH and 0.1% (w/v) polyvinyl alcohol in humidified atmosphere of 5% CO2 and 5% O2. The cortices were divided in six treatment groups: 1) 10 ng/mL EGF (EGF V0); 2) 10 ng/mL of EGF plus 1 mM Z-VAD-FMK (EGF V1); 3) 10 ng/mL of EGF and 10 mM Z-VAD-FMK (EGF V10); 4) 1 mM Z-VAD-FMK; 5) 10 mM Z-VAD-FMK and (6) no EGF and Z-VAD-FMK supplementation. Ovarian follicles within the tissues were processed for histology and assessed for follicle density, viability (based on morphology) and diameter immediately after collection (Control) or after 3 or 7 days of in vitro incubation. Comparison among fresh and culture treatment group was performed using ANOVA test. There were no differences (P > 0.05) in follicle density and viability among different culture treatments. However, there were differences in this parameter between culture days. Specifically, culturing tissue for 7 days resulted in significant reduction in follicle viability and density, regardless of treatments. We found a difference in size between culture days when these follicles were cultured using 10 mM Z-VAD-FMK or 10 ng/mL EGF (EGF V0). In sum, the finding demonstrated that Z-VAD-FMK at the dosage used in the present study does not provide the protective effect to ovarian tissue during in vitro culture. Future studies should explore different Z-VAD-FMK dosages or other anti-apoptotic agent, such as surviving in protecting ovarian follicles against cell death.

Keywords: anti apoptotic drug, bitches, follicles, Z-VAD-FMK

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Authors: Madhavi S. Apte, Milind Bhitre

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In pharmaceutical research and new drug development, medicinal plants have important roles. Similarly, Indian dhak tree belonging to family Fabaceae has been widely used in the traditional Indian medical system of ‘Ayurveda’ for the treatment of a variety of ailments. Hence the cell viability study was undertaken to evaluate and compare the activity of extracts of various parts like flower, bark, leaf, seed by conducting MTT assay method along with other pharmacognostical studies. The methanolic extracts of bark, flowers, leaves, and seeds were used for the study. The cell viability MTT assay was performed using the standard operating procedures. The extracts were dissolved in DMSO and serially diluted with complete medium to get the concentrations range of test concentration. DMSO concentration was kept < 0.1% in all the samples. HUVEC cells maintained in appropriate conditions were seeded in 96 well plates and treated with different concentrations of the test samples and incubated at 37°C, 5% CO₂ for 96 hours. MTT reagent was added to the wells and incubated for 4 hours; the dark blue formazan product formed by the cells was dissolved in DMSO under a safety cabinet and read at 550nm. Percentage inhibitions were calculated and plotted with the concentrations used to calculate the IC50 values. The bark, flower, leaves and seed extracts have shown the cytotoxicity activity and can be further studied for antiangiogenesis activity.

Keywords: pharmacognosy, Cell viability, MTT assay, anti-angiogenesis

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Authors: Andualem Workie

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In this study, we examined the sinterability and bioactivity of MgO-SiO₂-P₂O₅-CaO-CaF₂ glass compositions created through spray pyrolysis. We evaluated the bioactivity of the materials by immersing them for varying periods of time in simulated bodily fluid (SBF) and found that bioactivity was related to the sintering temperature and soaking time. The material's pH value during immersion in SBF was within the range of 7.4-8.2, which is below 8.5 and improves compatibility and reduces toxicity in biological applications. We used X-ray diffraction and scanning electron microscopy to determine the phase compositions and morphologies of the samples and found that the 1100°C sintered A-W GC sample exhibited the highest bioactivity after soaking in SBF. This sample was dominated by fluorapatite, wollastonite, and whitlockite crystals scattered throughout the glass matrix. The crystallinity (%) of the A-W GC increased as its bioactivity improved, making it more suitable for use in pharmaceutical applications. We also conducted a cytotoxicity test on A-W GC samples sintered at different temperatures and found that the glass-ceramics were non-toxic to MC3T3-E1 cells at all extraction concentrations, except for those sintered at 700°C at concentrations of 250, 200, and 150 mg/ml where cell viability (%) was below the threshold of 70%.

Keywords: apatite wollastonite glass ceramics, bioactivity, calcination, cell viability

Procedia PDF Downloads 63

Authors: S. Ahmed John

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Marine forms-bacteria, actinobacteria, cynobacteria, fungi, microalgae, seaweeds mangroves and other halophytes an extremely important oceanic resources and constituting over 90% of the oceanic biomass. The marine natural products have lead to the discovery of many compounds considered worthy for clinical applications. The marine sources have the highest probability of yielding natural products. Natural derivatives play an important role to prevent the cancer incidences as synthetic drug transformation in mangrove. 28.12% of anticancer compound extracted from the mangroves. Exchocaria agollocha has the anti cancer compounds. The present investigation reveals the potential of the Exchocaria agollocha with biotechnological applications for anti cancer, antimicrobial drug discovery, environmental remediation, and developing new resources for the industrial process. The anti-cancer activity of Exchocaria agollocha was screened from 3.906 to 1000 µg/ml of concentration with the dilution leads to 1:1 to 1:128 following methanol and chloroform extracts. The cell viability in the Exchocaria agollocha was maximum at the lower concentration where as low at the higher concentration of methanol and chloroform extracts when compare to control. At 3.906 concentration, 85.32 and 81.96 of cell viability was found at 1:128 dilution of methanol and chloroform extracts respectively. At the concentration of 31.25 following 1:16 dilution, the cell viability was 65.55 in methanol and 45.55 in chloroform extracts. However, at the higher concentration, the cell viability 22.35 and 8.12 was recorded in the extracts of methanol and chloroform. The cell viability was more in methanol when compare to chloroform extracts at lower concentration. The present findings gives current trends in screening and the activity analysis of metabolites from mangrove resources and to expose the models to bring a new sustain for tackling cancer. Bioactive compounds of Exchocaria agollocha have extensive use in treatment of many diseases and serve as a compound and templates for synthetic modification.

Keywords: bio-active product, compounds, natural products and microalgae

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Authors: Zenab Naseem, Sadia Imran

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One of the key solutions to the climate change crisis is to develop renewable energy resources, such as solar and wind power and biogas. This paper explores the socioeconomic and environmental viability of solar energy, based on a case study of the Soan Valley Development Program. Under this project, local farmers were provided solar water pumps at subsidized rates. These have been functional for the last seven years and have gained popularity among the local communities. The study measures the economic viability of using solar energy in agriculture, based on data from 36 households, of which 12 households each use diesel, electric and solar water pumps. Our findings are based on the net present value of each technology type. We also carry out a qualitative assessment of the social impact of solar water pumps relative to diesel and electric pumps. Finally, we conduct an environmental impact assessment, using the lifecycle assessment approach. All three analyses indicate that solar energy is a viable alternative to diesel and electricity.

Keywords: alternative energy sources, pollution control adoption and costs, solar energy pumps, sustainable development

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Authors: Abdurzag Kerban

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Preservation of semen had a major impact on sheep genetic breeding. The aim of this study was to evaluate the viability of ram spermatozoa after freezing pellet using cold surfaces made from cattle fat and paraffin wax. A pool of three to four ejaculates were pooled from six rams within a period of ten weeks. Semen was diluted in egg yolk-Tris diluent and processed in 0.25 ml straw and 0.1 ml pellets. Motility was evaluated after dilution, before freezing and post-thawing at 0, 1, 2 and 3 hour incubation. Viability index, acrosome integrity and leakage of intracellular enzymes (aspartat aminotransferase and alkline phosphatase) were also evaluated. Spermatozoa exhibited highly significant percentages of motility at 0, 1, 2 and 3 hours incubation after thawing and viability index in 0.25 ml straw and 0.1 ml pellets on cattle fat plate as compared to ram spermatozoa frozen on paraffin wax. In conclusion, cattle fat plate could be used as the cold surface of choice for freezing ram semen in form of pellets. Such form of frozen semen could be used as efficiently as semen frozen in straws. This simple method is economical with little expensive equipment or supplies, and may provide an efficient technique to cryopreserve ram spermatozoa in developing countries.

Keywords: ram semen, freezing, straw, pellet

Procedia PDF Downloads 550

Authors: Malik Shahzad Shabbir, Muhammad Saarim Ghazi, Amir Khalil ur Rehman

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This study examines the factors which influence and contribute towards the financial viability of Islamic finance and its impact on global financial stability. However, the purpose of this paper is to differentiate the practical implications of both Islamic and conventional finance on global financial stability. The Islamic finance is asset backed financing which creates wealth through trade, commerce and believes in risk and return sharing. Islamic banking is asset driven as against to conventional banking which is liability driven. In order to introduce new financial products for market, financial innovation in Islamic finance must be within the Shari’ah parameters that are tested against the ‘Maqasid al-Shari’ah’. Interest-based system leads to income and wealth inequalities and mis-allocation of resources. Moreover, this system has absence of just and equitable aspect of distribution that may exploit either the debt holder or the financier. Such implications are reached to a tipping point that leaves only one choice: change or face continued decline and misery.

Keywords: viability, global financial stability, practical implications, asset driven, tipping point

Procedia PDF Downloads 271

Authors: Muhammad Zubair, Maqbool Ahmad, Al-Hafizah Shafia Tehseen Gul, Shujait Ali

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The use of vitamins has significant effects on the reproductive system of mammals. The present study was conducted to investigate the useful effects of vitamin E on reproductive functions of Teddy bucks. For this purpose, 8 adult Teddy bucks were randomly divided into two treatment groups viz; A (control) and B (vitamin E with dose of 200 mg/kg BW/day). These treatments continued for 12 weeks. Semen quality parameters (volume, motility, sperm morphology and sperm DNA integrity) of experimental bucks of each group was evaluated on weekly basis, while testicular measurements (length, scrotal circumference and weights) were recorded at 0 and 12th week of experiment. Serum concentrations of male sex hormones (testosterone, LH, FSH) and cortisol were recorded fortnightly. Similarly, body weights of bucks were also measured fortnightly until completion of the study. The data were subjected to two-way analysis of variance, followed by Duncan test for multiple mean comparisons. Supplementation of vitamin E improved significantly (P<0.05) the semen quality parameter, body weights, testicular measurements and serum levels of sex hormones. However, there was no effect on serum cortisol. It was concluded from the present study that dietary supplementation of vitamin E has beneficial effects on the semen and hormones in male reproductive system.

Keywords: hormones, semen, teddy bucks, testicular measurements

Procedia PDF Downloads 265

Authors: Dong Xie, Leah Howard, Yiming Weng

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The objective of this study was to synthesize and characterize 5-acryloyloxy-3,4-dichlorocrotonolactone (a furanone derivative), use this derivative to modify a dental restorative, and study the effect of the derivative on the antibacterial activity and compressive strength of the formed restorative. In this study, a furanone derivative was synthesized, characterized, and used to formulate a dental restorative. Compressive strength (CS) and S. mutans viability were used to evaluate the mechanical strength and antibacterial activity of the formed restorative. The fabricated restorative specimens were photocured and conditioned in distilled water at 37oC for 24 h, followed by direct testing for CS or/and incubating with S. mutans for 48 h for antibacterial testing. The results show that the modified dental restorative showed a significant antibacterial activity without substantially decreasing the mechanical strengths. With addition of the antibacterial derivative up to 30%, the restorative kept its original CS nearly unchanged but showed a significant antibacterial activity with 68% reduction in the S. mutans viability. Furthermore, the antibacterial function of the modified restorative was not affected by human saliva. The aging study also indicates that the modified restorative may have a long-lasting antibacterial function. It is concluded that this experimental antibacterial restorative may potentially be developed into a clinically attractive dental filling restorative due to its high mechanical strength and antibacterial function.

Keywords: antibacterial, dental restorative, compressive strength, S. mutans viability

Procedia PDF Downloads 296

Authors: Lourdita Llanto

Abstract:

This study described and assessed the implementation of artificial insemination (AI) for cattle and carabaos in the Bicol Region, Philippines: Albay, Sorsogon and Camarines Sur. Three hundred respondents were interviewed. Results were analyzed using frequency counts, means, percentages and chi-square test. Semen samples from different stations were analyzed for motility, viability and morphology. T-test was used in semen quality evaluation. Provincial AI coordinators (PAIC) were male, averaging 59 years old, married, had college education, served in government service for 34 years, but as PAIC for 5.7 years. All had other designations. Mean AI operation was 11.33 years with annual support from the local government unit of Php76,666.67. AI technicians were males, married, with college education, and trained on AI. Problems were on mobility; inadequate knowledge of farmers in animal raising and AI; and lack of liquid nitrogen and frozen semen supply. There was 2.95 municipalities and breedable cattle/carabaos of 3,091.25 per AI technician. Mean number of artificially inseminated animals per AI technician for 2011 was 28.57 heads for carabaos and 8.64 heads for cattle. There was very low participation rate among farmers. Carabaos were 6.52 years with parity 1.53. Cattle were 5.61 years, with parity of 1.51. Semen quality significantly (p ≤ 0.05) deteriorated in normal and live sperm with storage and handling at the provincial and field stations. Breed, AI technicians practices and AI operation significantly affected conception rate. Mean conception rate was 57.62%.

Keywords: artificial insemination, carabao, parity, mother tanks, frozen semen

Procedia PDF Downloads 414