Total Incubation Time Research Articles

X-ray crystal structure of proliferating cell nuclear antigen 1 from Aeropyrum pernix.

Proliferating cell nuclear antigen (PCNA) plays a critical role in DNA replication by enhancing the activity of various proteins involved in replication. Inthis study, the crystal structure of ApePCNA1, one of three PCNAs from thethermophilic archaeon Aeropyrum pernix, was elucidated. ApePCNA1 was cloned and expressed in Escherichia coli and the protein was purified and crystallized. The resulting crystal structure determined at 2.00 Å resolution revealed that ApePCNA1 does not form a trimeric ring, unlike PCNAs from other domains of life. It has unique structural features, including a long interdomain-connecting loop and a PIP-box-like sequence at the N-terminus, indicating potential interactions with other proteins. These findings provide insights into the functional mechanisms of PCNAs in archaea and their evolutionary conservation across different domains of life. A modified medium and protocol were used to express recombinant protein containing the lac operon. The expression of the target protein increased and the total incubation time decreased when using this system compared with those of previous expression protocols.

Eco-friendly Biosynthesis of Silver Nanoparticles from Aloe vera Leaves Based on Various Parameters

In the current study, an ecofriendly and economically very effective method for the biosynthesis of silver nanoparticles (AgNPs) using aqueous extract of Aloe vera leaves has been investigated. The synthesis of AgNPs was checked by the change of color of the solution from yellow to dark blue or black due to a phenomenon known as surface plasmon resonance. The ability of silver nanoparticles to get effected by various process parameters like volume of Aloe vera leaves extract, concentration of silver nitrate solution, and the total incubation time was checked. The analysis and characterization was done using UV-Visible spectroscopy and SEM-EDX. The SEM-EDX analysis confirmed the size, shape and formation of the silver nanoparticles. The best conditions needed for the synthesis of AgNPs using Aloe vera leaves extract was found to be 5mM concentration of silver nitrate solution using 15% volume of Aloe vera leaves extract and incubated for 10minutes under the sunlight.

Combination of the electronic nose with microbiology as a tool for rapid detection of Salmonella

Salmonella is one of the most important foodborne pathogens and its analysis in raw and processed products is mandatory in the food industry. Although microbiological analysis is the standard practice for Salmonella determination, these assays are commonly laborious and time-consuming, thus, alternative techniques based on easy operation, few manipulation steps, low cost, and reduced time are desirable. In this paper, we demonstrate the use of an e-nose based on ionogel composites (ionic liquid + gelatine + Fe3O4 particles) as a complementary tool for the conventional microbiological detection of Salmonella. We used the proposed methodology for differentiating Salmonella from Escherichia coli, Pseudomonas fluorescens, Pseudomonas aeruginosa, and Staphylococcus aureus in nonselective medium: pre-enrichment in brain heart infusion (BHI) (incubation at 35 °C, 24 h) and enrichment in tryptone soy agar (TSA) (incubation at 35 °C, 24 h), whereas Salmonella differentiation from E. coli and P. fluorescens was also evaluated in selective media, bismuth sulfite agar (BSA), xylose lysine deoxycholate agar (XLD), and brilliant green agar (BGA) (incubation at 35 °C, 24 h). The obtained data were compared by principal component analysis (PCA) and different machine learning algorithms: multilayer perceptron (MLP), linear discriminant analysis (LDA), instance-based (IBk), and Logistic Model Trees (LMT). For the nonselective media, under optimized conditions, taking merged data of BHI + TSA (total incubation time of 48 h), an accuracy of 85% was obtained with MLP, LDA, and LMT, while five separated clusters were presented in PCA, each cluster corresponding to a bacterium. In addition, for evaluation of the e-nose for discrimination of Salmonella using selective media, considering the combination of BSA + XLD and total incubation of 72 h, the PCA showed three separated and well-defined clusters corresponding to Salmonella, E. coli, and P. fluorescens, and an accuracy of 100% was obtained for all classifiers.

Dual inhibition of complement C5 and CD14 attenuates inflammation in a cord blood model.

Escherichia coli and Group B streptococci (GBS) are the main causes of neonatal early-onset sepsis (EOS). Despite antibiotic therapy, EOS is associated with high morbidity and mortality. Dual inhibition of complement C5 and the Toll-like receptor co-factor CD14 has in animal studies been a promising novel therapy for sepsis. Whole blood was collected from the umbilical cord after caesarean section (n = 30). Blood was anti-coagulated with lepirudin. C5 inhibitor (eculizumab) and anti-CD14 was added 8 min prior to, or 15 and 30 min after adding E. coli or GBS. Total bacterial incubation time was 120 min (n = 16) and 240 min (n = 14). Cytokines and the terminal complement complex (TCC) were measured using multiplex technology and ELISA. Dual inhibition significantly attenuated TCC formation by 25-79% when adding inhibitors with up to 30 min delay in both E. coli- and GBS-induced inflammation. TNF, IL-6 and IL-8 plasma concentration were significantly reduced by 28-87% in E. coli-induced inflammation when adding inhibitors with up to 30 min delay. The dual inhibition did not significantly reduce TNF, IL-6 and IL-8 plasma concentration in GBS-induced inflammation. Dual inhibition of C5 and CD14 holds promise as a potential future treatment for severe neonatal EOS. Neonatal sepsis can cause severe host inflammation with high morbidity and mortality, but there are still no effective adjunctive immunologic interventions available. Adding CD14 and complement C5 inhibitors up to 30 min after incubation of E. coli or Group B streptococci in a human umbilical cord blood model significantly reduced complement activation and cytokine release. Dual inhibition of C5 and CD14 is a potential future therapy to modulate systemic inflammation in severe cases of neonatal sepsis.

Attomolar sensitivity microRNA detection using real-time digital microarrays

MicroRNAs (miRNAs) are a family of noncoding, functional RNAs. With recent developments in molecular biology, miRNA detection has attracted significant interest, as hundreds of miRNAs and their expression levels have shown to be linked to various diseases such as infections, cardiovascular disorders and cancers. A powerful and high throughput tool for nucleic acid detection is the DNA microarray technology. However, conventional methods do not meet the demands in sensitivity and specificity, presenting significant challenges for the adaptation of miRNA detection for diagnostic applications. In this study, we developed a highly sensitive and multiplexed digital microarray using plasmonic gold nanorods as labels. For proof of concept studies, we conducted experiments with two miRNAs, miRNA-451a (miR-451) and miRNA-223-3p (miR-223). We demonstrated improvements in sensitivity in comparison to traditional end-point assays that employ capture on solid phase support, by implementing real-time tracking of the target molecules on the sensor surface. Particle tracking overcomes the sensitivity limitations for detection of low-abundance biomarkers in the presence of low-affinity but high-abundance background molecules, where endpoint assays fall short. The absolute lowest measured concentration was 100 aM. The measured detection limit being well above the blank samples, we performed theoretical calculations for an extrapolated limit of detection (LOD). The dynamic tracking improved the extrapolated LODs from femtomolar range to sim 10 attomolar (less than 1300 copies in 0.2 ml of sample) for both miRNAs and the total incubation time was decreased from 5 h to 35 min.

Optimization of the Transformation Protocol for Increased Efficiency of Genetic Transformation in Hevea brasiliensis.

The recurring growth of bacterium in newly developed resistant cells and a minimal level of bacterial infection rate are the main limiting factors of Agrobacterium-mediated transformation experiments in Hevea brasiliensis. The current study aimed to optimize crucial factors of the transformation protocol in order to obtain an efficient transformation experimental model for Hevea using cotyledonary somatic embryos as explants. Transformation conditions such as antibiotic concentration, preculture duration, Agrobacterium concentration, sonication and cocultivation conditions were analyzed using the binary vector pCAMBIA2301. Transient transformation was confirmed by GUS histochemical staining. The best transformation efficiency was observed when the explants were not cultured on a preculture medium that contained acetosyringone at a level of 100 μM. The best results were obtained using a bacterial density of 0.45 at OD 600 nm, 50 s of sonication of explants in a bacterial liquid culture and a total incubation time of 18 min in the same bacterial suspension. Transmission electron microscopical analysis confirmed the impacts of sonication on bacterial infection efficiency. Cocultivation conditions of 22 °C and 84 h of darkness were optimal for the transfer of T-DNA. Agrobacterium was eliminated with 500 mg/L of timentin, and the selection of transformants was performed using 100 mg/L of kanamycin in the selection medium. The presence of transgene was confirmed in the resistant embryos by polymerase chain reaction (PCR). The improved method of genetic transformation established in the present study will be useful for the introduction of foreign genes of interest into the Hevea genome for the breeding of this economically important plant species in the future.

Incubation temperature and physiological aging in the zebra finch.

In birds, incubation temperature has received increased attention as an important source of phenotypic variability in offspring. A lower than optimal incubation temperature may negatively affect aspects of nestling physiology, such as body growth and energy metabolism. However, the long-term effects of sub-optimal incubation temperature on morphology and physiology are not well understood. In a previous study, we showed that zebra finches from eggs incubated at a low temperature (35.9°C) for 2/3 of the total incubation time suffered a lower post-fledging survival compared to individuals that had been incubated at higher temperatures (37.0 and 37.9°C). In the present study, we investigated whether these variations in incubation temperature could cause permanent long-lasting differences in body mass, body size, or basal metabolic rate. Furthermore, we tested whether the observed differences in survival between treatment groups would be reflected in the rate of physiological deterioration, assessed through oxidative damage and decreased metabolic rate with age (i.e. ‘metabolic aging’). Incubation temperature did not significantly affect embryonic or nestling body growth and did not influence final adult body mass or body size. Nor was there any long-term effect on basal metabolic rate. Birds from eggs incubated at the lowest temperature experienced an accumulation of oxidative damage with age, although this was not accompanied by an accelerated rate of metabolic aging. The present results suggest that the low survival in these birds was possibly mediated by increased oxidative stress, but independent of body growth and the basal metabolic rate.

Optimization of Biomethane Production via Fermentation of Chicken Manure Using Marine Sediment: A Modeling Approach Using Response Surface Methodology

In this study, marine sediment (MS) was successfully used as a source of methanogenic bacteria for the anaerobic digestion (AD) of chicken manure (CM). Using MS showed high production in liquid and semi-solid conditions. Even in solid conditions, 169.3 mL/g volatile solids of chicken manure (VS-CM) was produced, despite the accumulation of ammonia (4.2 g NH3-N/kg CM). To the best of our knowledge, this is the highest methane production from CM alone, without pretreatment, in solid conditions (20%). Comparing MS to Ozouh sludge (excess activated sewage sludge) (OS), using OS under semi-solid conditions resulted in higher methane production, while using MS resulted in more ammonia tolerance (301 mL/gVS-CM at 8.58 g NH3-N/kg). Production optimization was carried out via a response surface methodology (RDM) model involving four independent variables (inoculum ratio, total solid content, NaCl concentration, and incubation time). Optimized methane production (324.36 mL/gVS-CM) was at a CM:MS ratio of 1:2.5 with no NaCl supplementation, 10% total solid content, and an incubation time of 45 days.

Physicochemical and gelling properties of whey protein hydrolysates generated at 5 and 50 °C using Alcalase® and Neutrase®, effect of total solids and incubation time

Gelation temperature (Tg), apparent viscosity (ηapp), turbidity and chromatography profiles of whey protein concentrate (WPC) and corresponding hydrolysates (WPHs) generated with Alcalase® and Neutrase® at different temperatures and total solids (TS) were compared. WPC incubated with Alcalase at 50 °C exhibited the highest rate and degree of hydrolysis (DH). After 4 h, the 5 °C Alcalase WPH (10% TS) had a DH and molecular mass distribution similar to those generated at 5 and 50 °C with Neutrase; however, the chromatography profiles differed. Tg was lower for WPHs than for WPC and depended on both the enzyme and TS. Moreover, the Tgs were lower for the Alcalase hydrolysates than for those generated with Neutrase. The ηapp and turbidity properties of the hydrolysates depended on the enzyme used, solution TS and incubation temperature. Hydrolysates had lower ηapp than WPC. The 50 °C Neutrase hydrolysates displayed lower turbidity than the corresponding Alcalase hydrolysates.

Ontogeny of the Moorish gecko Tarentola mauritanica with emphasis on morphogenesis of the skin and its derivatives.

Studying reptilian embryonic development provides answers to many questions related to the development of tetrapods. Reptilian skin has been recently considered in studies at the evo-devo level. The lizard epidermis has to be shed periodically. At the embryonic level, contention exists regarding the first layers to appear, whether the oberhautchen or the clear layer, and whether the shedding complex develops before hatching. Geckos exhibit diverse morphologies independently evolved multiple times within the clade, such as subdigital pad lamellae. Here we investigate the embryonic development of Tarentola mauritanica and establishing its embryonic table. Primarily we follow the development of the integument. This is a closely related species to Tarentola annularis and it is crucial to investigate whether it has the same derived digital condition of claw regression. Eleven embryonic stages are described according to the external morphological characteristics of the embryos. Interestingly, the oviposition stage appears earlier than its close relative T. annularis, and the total incubation time is less. We also describe skin development, adding clear evidence to the debate on the development of the shedding complex, which we found is developed before hatching. We describe one layer of periderm and the clear layer as the first embryonic epidermal layers. Generally, our results show the genus Tarentola to have the advantage of being a unique taxon, easily breed at the laboratory, with multiple clutches per year, and with an earlier stage at oviposition. That could be a model animal for embryonic development and experimental embryology studies.

Biostimulation enhanced the biotic degradation of hexabromocyclododecane in sediments

The aim of this study was to investigate biodegradation of γ-hexabromocyclododecane (γ-HBCDD) under conditions mimicking three bioremediation strategies: (i) biostimulation: addition of sodium formate and ethanol to stimulate biodegradation as the carbon source and electron donor, respectively; (ii) bioaugmentation: addition of an enrichment culture of Dehalobium chlorocoercia strain DF-1; and (iii) natural attenuation: no amendments. To differentiate between biotic and abiotic mechanisms affecting γ-HBCDD degradation, four control microcosms were set up as sterile, negative, abiotic, and contaminant control. Sediment microcosms were prepared in 20-mL bottles and operated as duplicate sacrificial reactors with a sediment-to-liquid ratio of 3 g wet solid:3.5 mL liquid. Total incubation time was 36 days with sampling every 4 days, except the last day. γ-HBCDD contents of sediments were extracted using ultrasonication and analyzed using GC-MS. Four control microcosms were used to observe the effect of (i) microbial activity (sterilization with mercuric chloride and autoclaving), i.e., sterile; (ii) microbial culture without DF-1 cells, i.e., negative control; (iii) sediments, where kaolinite is used instead of sediments, i.e., abiotic control; and (iv) γ-HBCDD, where no analyte is added, i.e., contaminant control. Biostimulation showed the highest γ-HBCDD biodegradation rate (k = 0.0542 day−1) and enhanced biodegradation compared to natural attenuation (k = 0.0155 day−1). Bioaugmentation (k = 0.0123 day−1) with DF-1 strain showed a sharp decrease at the beginning, but could not maintain this trend afterwards. Paired comparison of microcosms yielded no statistically significant difference between bioaugmentation and natural attenuation; hence, DF-1 strain did not improve degradation when compared to natural attenuation. This was also substantiated by observations from the negative control set. Sterile and abiotic control sets showed no significant concentration change in time. Consequently, adsorption was not considered as a significant mechanism acting on γ-HBCDD concentration change in our sediment microcosms. Thus, γ-HBCDD decrease observed in bioremediation microcosms was attributed to microbial activity. We reported effective analyte degradation with biostimulation. This was the first study to test bioaugmentation for HBCDD degradation, but we observed no enhancement of degradation with the DF-1 strain tested. Previous studies observed HBCDD reduction in their sterilized controls, hence reported total biotic and abiotic degradation rate. In this study, comparative evaluation of three test and four control microcosms enabled identification of only anaerobic biodegradation rates for γ-HBCDD, providing useful information for bioremediation of contaminated sites.

Performance Comparison of Francis Media with Other Methods in the Identification of Burkholderia pseudomallei

Burkholderia pseudomallei is the causative agent for melioidosis, severe infection that is associated with high morbidity and mortality. Difficulties in laboratory diagnosis of melioidosis may delay the treatment and as a result, affect disease outcomes. Culture is the mainstay for laboratory diagnosis of the disease followed by further commercial biochemical testing for identification of organism. The whole process may take up to 72 hours. Furthermore, commercial identification system may fail to distinguish between B. pseudomallei and closely related species like B. cepacia. The objective of this study was to compare the efficacy of Francis media against commercial identification system such as API20NE and VITEK2GN for the identification of B. pseudomallei. This study showed that Francis media was 100% reliable in the identification of B. pseudomallei as compared to others. The media also offered fast identification of organism from clinical specimen with total incubation time required of only 18 hours. API20NE was able to correctly identify only 5 (12.5%) within 24 hours and 31 (78%) within 48 hours of incubation. VITEK2GN, identified 95% of the isolates correctly within 9 hours of incubation. However, both methods required an additional sub culturing prior to testing. Molecular method was used as the confirmatory method for identification of the isolate.

Characterization of Plasmodium Lactate Dehydrogenase and Histidine-Rich Protein 2 Clearance Patterns via Rapid On-Bead Detection from a Single Dried Blood Spot.

.A rapid, on-bead enzyme-linked immunosorbent assay for Plasmodium lactate dehydrogenase (pLDH) and Plasmodium falciparum histidine-rich protein 2 (HRP2) was adapted for use with dried blood spot (DBS) samples. This assay detected both biomarkers from a single DBS sample with only 45 minutes of total incubation time and detection limits of 600 ± 500 pM (pLDH) and 69 ± 30 pM (HRP2), corresponding to 150 and 24 parasites/μL, respectively. This sensitive and reproducible on-bead detection method was used to quantify pLDH and HRP2 in patient DBS samples from rural Zambia collected at multiple time points after treatment. Biomarker clearance patterns relative to parasite clearance were determined; pLDH clearance followed closely with parasite clearance, whereas most patients maintained detectable levels of HRP2 for 35–52 days after treatment. Furthermore, weak-to-moderate correlations between biomarker concentration and parasite densities were found for both biomarkers. This work demonstrates the utility of the developed assay for epidemiological study and surveillance of malaria.

Sediment nutrient drivers of the growth dynamics of the rare fern Marsilea quadrifolia

In analysing the global decline of macrophytes, a scant attention was generally paid to the direct relations of rooted aquatic plants with sediment eutrophication. Nevertheless, the sediment is a pivotal driver of macrophyte dynamics. In order to add new data about aquatic plant–sediment interactions, a microcosm experiment was performed to gain knowledge on the growth dynamics of the rare aquatic fern Marsilea quadrifolia L. Healthy shoots of M. quadrifolia were grown under an increasing availability of nutrients (N and P) and reducing sediment conditions for a total incubation time of 42 days. The variation in biomass and number of fronds and knots was monitored regularly at intervals of 10–11 days. The results confirm the pivotal role of sediment in driving the growth of M. quadrifolia, which responds rapidly to the increase in nutrient availability up to a moderate excess. M. quadrifolia seems to be a slightly eutrophic-rooted aquatic species, able to tolerate pore-water concentrations up to ~450 and 4 μM of NH4 + and PO4 3−, respectively. The present data corroborate the need to consider the short-term dynamics when analysing plant adaptations to sediment eutrophication, as well as a physical and chemical characterization of sediments to evaluate their potential bioreceptivity.

Simultaneous capture and sequential detection of two malarial biomarkers on magnetic microparticles

We have developed a rapid magnetic microparticle-based detection strategy for malarial biomarkers Plasmodium lactate dehydrogenase (pLDH) and Plasmodium falciparum histidine-rich protein II (PfHRPII). In this assay, magnetic particles functionalized with antibodies specific for pLDH and PfHRPII as well as detection antibodies with distinct enzymes for each biomarker are added to parasitized lysed blood samples. Sandwich complexes for pLDH and PfHRPII form on the surface of the magnetic beads, which are washed and sequentially re-suspended in detection enzyme substrate for each antigen. The developed simultaneous capture and sequential detection (SCSD) assay detects both biomarkers in samples as low as 2.0parasites/µl, an order of magnitude below commercially available ELISA kits, has a total incubation time of 35min, and was found to be reproducible between users over time. This assay provides a simple and efficient alternative to traditional 96-well plate ELISAs, which take 5–8h to complete and are limited to one analyte. Further, the modularity of the magnetic bead-based SCSD ELISA format could serve as a platform for application to other diseases for which multi-biomarker detection is advantageous.

Improved standardization and potential for shortened time to results with BD Kiestra™ total laboratory automation of early urine cultures: A prospective comparison with manual processing

We compared the results of 505 urine specimens prospectively processed by both conventional manual processing (MP) with 16–24h incubation to BD Kiestra™ Total Laboratory Automation (TLA) system with a shortened incubation of 14h: 97% of culture results were clinically concordant. TLA processing was associated with improved standardization of time of first culture reading and total incubation time.

High atmospheric temperatures and 'ambient incubation' drive embryonic development and lead to earlier hatching in a passerine bird.

Tropical and subtropical species typically experience relatively high atmospheric temperatures during reproduction, and are subject to climate-related challenges that are largely unexplored, relative to more extensive work conducted in temperate regions. We studied the effects of high atmospheric and nest temperatures during reproduction in the zebra finch. We characterized the temperature within nests in a subtropical population of this species in relation to atmospheric temperature. Temperatures within nests frequently exceeded the level at which embryo’s develop optimally, even in the absence of parental incubation. We experimentally manipulated internal nest temperature to demonstrate that an average difference of 6°C in the nest temperature during the laying period reduced hatching time by an average of 3% of the total incubation time, owing to ‘ambient incubation’. Given the avian constraint of laying a single egg per day, the first eggs of a clutch are subject to prolonged effects of nest temperature relative to later laid eggs, potentially increasing hatching asynchrony. While birds may ameliorate the negative effects of ambient incubation on embryonic development by varying the location and design of their nests, high atmospheric temperatures are likely to constitute an important selective force on avian reproductive behaviour and physiology in subtropical and tropical regions, particularly in the light of predicted climate change that in many areas is leading to a higher frequency of hot days during the periods when birds breed.

Role of biotransformation, sorption and mineralization of 14C-labelled sulfamethoxazole under different redox conditions

14C-sulfamethoxazole biotransformation, sorption and mineralization was studied with heterotrophic and autotrophic biomass under aerobic and anoxic conditions, as well as with anaerobic biomass. The 14C-radiolabelled residues distribution in the solid, liquid and gas phases was closely monitored along a total incubation time of 190h. Biotransformation was the main removal mechanism, mineralization and sorption remaining below 5% in all the cases, although the presence of a carbon source exerted a positive effect on the mineralization rate by the aerobic heterotrophic bacteria. In fact, an influence of the type of primary substrate and the redox potential was observed in all cases on the biotransformation and mineralization rates, since an enhancement of the removal rate was observed when an external carbon source was used as a primary substrate under aerobic conditions, while a negligible effect was observed under nitrifying conditions. In the liquid phases collected from all assays, up to three additional peaks corresponding to 14C-radiolabelled residues were detected. The highest concentration was observed under anaerobic conditions, where two radioactive metabolites were detected representing each around 15% of the total applied radioactivity after 180h incubation. One of the metabolites detected under anoxic and anaerobic conditions, is probably resulting from ring cleavage of the isoxazole ring.

Optimization, production and scale up of debittered kinnow beverage by and #945;-L-rhamnosidase producing yeast

α-L-Rhamnosidase is used for debittering the citrus juice by hydrolyzing bitter naringin to nonbitter prunin and rhamnose. The present study was carried out on fermentative production of debittered kinnow (Citrus reticulata Blanco) beverage using α-L-rhamnosidase producing yeast in order to utilize its immense potentiality in processed kinnow juice industry. The effect of percent yeast inoculum concentration (0.25-1.25 v/v), total soluble solids (12-16 °B), temperature (15-35 °C) and incubation time (12-60 h) were studied to optimize the production of α-L- rhamnosidase enzyme from Clavispora lusitaniae in kinnow juice. Results indicated that yeast showing maximum rhamnosidase activity (0.056 IU mL-1) in presence of yeast inoculum concentration (0.75% v/v), brix (13 °B), temperature (30±5 °C) and incubation time (48 h). Further, these optimized conditions were used in upscale production of debittered kinnow beverage. The physicochemical parameters of freshly prepared beverage TSS 13.00±0.20 °B, acidity 0.14±0.03%, pH 3.40±0.10, brix acid ratio 92.85±0.00, limonin 6.90±0.10 ppm, naringin 443.00±10.00 ppm, total sugars 12.90±0.30%, reducing sugars 2.42±0.20 and ascorbic acid 27.80±1.00 mg/100 mL. All the physicochemical parameters did not change significantly during storage. The decrease of naringin with storage was 443.00±10.00 to 143.70±4.00 ppm due to the α-L-rhamnosidase activity of yeast. The percentage of ethanol and CO2 were 0.89±0.05% and 1.46±0.06 bar after three months of storage. All the sensory parameters like taste, color, aroma, bouquet, flavor and astringency of kinnow beverage were stable at storage period of 90 days with almost no change in organoleptic sensation. Thus the technology presented here, a very less time consuming and safe for production of debittered kinnow beverage on large scale in citrus food industries.

O-032 Serotonin Is A Selective Vasoconstrictor Of Chicken Embryo Ductus Arteriosus

Introduction Decreased platelet number and/or function have been related to patent ductus arterious (DA). Activated platelets release vasoactive products, including serotonin (5-HT) that might be relevant for DA homeostasis DA. The chicken embryo has emerged as a suitable model for the study of DA vascular biology. In the present study, we investigated the possible vasoactive role of 5-HT in the chicken DA. Methods Rings of the DA of 15- to 20-d-old chicken embryos (total incubation time 21-d) were studied in a wire myograph. The response to 5-HT was investigated under different O 2 tensions (3, 7, and 74 kPa). The responses to the 5-HT 1B/D receptor agonist sumatriptan, the 5-HT 2A/B/C receptor agonist DOI and the selective serotonin reuptake inhibitors fluoxetine and sertraline were also investigated. Results 5-HT (10 nM-0.3 mM) contracted the pulmonary side of the DA (PulmDA) in a concentration-dependent manner. By contrast, 5-HT induced negligible contractions in the vessels that surround the PulmDA (i.e., the pre- and post-ductal pulmonary arteries, and the aortic side of the DA). 5-HT-induced contraction increased with development (15-d >17-d >19-d=20-d). O 2 tension did not affect 5-HT-induced contraction but elimination of extracellular calcium completely abolished it. Sumatriptan and DOI also contracted the PulmDA in a concentration-dependent manner. By contrast, fluoxetine and sertraline evoked contractions at very high concentrations (>0.1 mM). Conclusions Our data indicate that 5-HT receptors are functionally present in the chicken DA and suggest that platelet-derived 5-HT may play a pivotal role in the postnatal closure of the DA.