Abstract: In this study, the mechanical properties of alginate hydrogel material for self-standing 3D scaffold architecture with proper shape fidelity are investigated. In-lab built 3D bio-printer extrusion-based technology is utilized to fabricate 3D alginate scaffold constructs. The pressure, needle speed and stage speed are varied using a computer-controlled system. The experimental result indicates that the concentration of alginate solution, calcium chloride (CaCl2) cross-linking concentration and cross-linking ratios lead to the formation of alginate hydrogel with various gelation states. Besides, the gelling conditions, such as cross-linking reaction time and temperature also have a significant effect on the mechanical properties of alginate hydrogel. Various experimental tests such as the material gelation, the material spreading and the printability test for filament collapse as well as the swelling test were conducted to evaluate the fabricated 3D scaffold constructs. The result indicates that the fabricated 3D scaffold from composition of 3.5% wt alginate solution, that is prepared in DI water and 1% wt CaCl2 solution with cross-linking ratios of 7:3 show good printability and sustain good shape fidelity for more than 20 days, compared to alginate hydrogel that is prepared in a phosphate buffered saline (PBS). The fabricated self-standing 3D scaffold constructs measured 30 mm × 30 mm and consisted of 4 layers (n = 4) show good pore geometry and clear grid structure after printing. In addition, the percentage change of swelling degree exhibits high swelling capability with respect to time. The swelling test shows that the geometry of 3D alginate-scaffold construct and of the macro-pore are rarely changed, which indicates the capability of holding the shape fidelity during the incubation period. This study demonstrated that the mechanical and physical properties of alginate hydrogel could be tuned for a 3D bio-printing extrusion-based system to fabricate self-standing 3D scaffold soft structures. This 3D bioengineered scaffold provides a natural microenvironment present in the extracellular matrix of the tissue, which could be seeded with the biological cells to generate the desired 3D live tissue model for in vitro and in vivo tissue engineering applications.
Abstract: Background: An outbreak of gastroenteritis occurred in December 2019 after students of a Senior High School in Accra were served with kenkey and fish during their dinner. An investigation was conducted to characterize the affected people, the source of contamination, the etiologic food and agent. Methods: An epidemiological study was conducted with cases selected from the student population who were ill. Controls were selected from among students who also ate from the school canteen during dinner but were not ill. Food history of each case and control was taken to assess their exposure status. Epi Info 7 was used to analyze the data obtained from the outbreak. Attack rates and odds ratios were calculated to determine the risk of foodborne infection for each of the foods consumed by the population. The source of contamination of the foods was ascertained by conducting an environmental risk assessment at the school. Results: Data were obtained from 126 students, out of which 57 (45.2%) were cases and 69 (54.8%) were controls. The cases presented with symptoms such as diarrhea (85.96%), abdominal cramps (66.67%), vomiting (50.88%), headache (21.05%), fever (17.86%) and nausea (3.51%). The peak incubation period was 18 hours with a minimum and maximum incubation periods of 6 and 50 hours respectively. From the incubation period, duration of illness and the symptoms, non-typhoidal salmonellosis was suspected. Multivariate analysis indicated that the illness was associated with the consumption of the fried fish served, however this was statistically insignificant (AOR 3.1.00, P = 0.159). No stool, blood or food samples were available for organism isolation and confirmation of suspected etiologic agent. The environmental risk assessment indicated poor hand washing practices on the part of both the food handlers and students. Conclusion: The outbreak could probably be due to the consumption of the fried fish that might have been contaminated with Salmonella sp. as a result of poor hand washing practices in the school.
Abstract: Anaerobic batch experiments were conducted to investigate the effect of magnetite-supplementation (7 mM) on methane production from digested sludge undergoing two different microbial growth phases, namely fresh sludge (exponential growth phase) and degassed sludge (endogenous decay phase). Three different particle sizes were assessed: small (50 - 150 nm), medium (168 – 490 nm) and large (800 nm - 4.5 µm) particles. Results show that, in the case of the fresh sludge, magnetite significantly enhanced the methane production rate (up to 32%) and reduced the lag phase (by 15% - 41%) as compared to the control, regardless of the particle size used. However, the cumulative methane produced at the end of the incubation was comparable in all treatment and control bottles. In the case of the degassed sludge, only the medium-sized magnetite particles increased significantly the methane production rate (12% higher) as compared to the control. Small and large particles had little effect on the methane production rate but did result in an extended lag phase which led to significantly lower cumulative methane production at the end of the incubation period. These results suggest that magnetite produces a clear and positive effect on methane production only when an active and balanced microbial community is present in the anaerobic digester. It is concluded that, (i) the effect of magnetite particle size on increasing the methane production rate and reducing lag phase duration is strongly influenced by the initial metabolic state of the microbial consortium, and (ii) the particle size would positively affect the methane production if it is provided within the nanometer size range.
Abstract: The incubation period is defined as the time from infection with a microorganism to development of symptoms. In this research, two disease models: one with incubation period and another without incubation period were studied. The study involves the use of a mathematical model with a single incubation period. The test for the existence and stability of the disease free and the endemic equilibrium states for both models were carried out. The fourth order Runge-Kutta method was used to solve both models numerically. Finally, a computer program in MATLAB was developed to run the numerical experiments. From the results, we are able to show that the endemic equilibrium state of the model with incubation period is locally asymptotically stable whereas the endemic equilibrium state of the model without incubation period is unstable under certain conditions on the given model parameters. It was also established that the disease free equilibrium states of the model with and without incubation period are locally asymptotically stable. Furthermore, results from numerical experiments using empirical data obtained from Nigeria Centre for Disease Control (NCDC) showed that the overall population of the infected people for the model with incubation period is higher than that without incubation period. We also established from the results obtained that as the transmission rate from susceptible to infected population increases, the peak values of the infected population for the model with incubation period decrease and are always less than those for the model without incubation period.
Abstract: Introduction: The application of fissure sealants is considered to be an important primary prevention method used in dental medicine. However, the formation of microleakage gaps between tooth enamel and the fissure sealant applied is one of the most common reasons of dental caries development in teeth with fissure sealants. The association between various dental biomaterials may limit the major disadvantages and limitations of biomaterials functioning in a complementary manner. The present study consists in the incorporation of a cariostatic agent – silver diamine fluoride (SDF) – in a resin-based fissure sealant followed by the study of release kinetics by spectrophotometry analysis of the association between both biomaterials and assessment of the inhibitory effect on the growth of the reference bacterial strain Streptococcus mutans (S. mutans) in an in vitro study. Materials and Methods: An experimental in vitro study was designed consisting in the entrapment of SDF (Cariestop® 12% and 30%) into a commercially available fissure sealant (Fissurit®), by photopolymerization and photocrosslinking. The same sealant, without SDF was used as a negative control. The effect of the sealants on the growth of S. mutans was determined by the presence of bacterial inhibitory halos in the cultures at the end of the incubation period. In order to confirm the absence of bacteria in the surface of the materials, Scanning Electron Microscopy (SEM) characterization was performed. Also, to analyze the release profile of SDF along time, spectrophotometry technique was applied. Results: The obtained results indicate that the association of SDF to a resin-based fissure sealant may be able to increase the inhibition of S. mutans growth. However, no SDF release was noticed during the in vitro release studies and no statistical significant difference was verified when comparing the inhibitory halo sizes obtained for test and control group. Conclusions: In this study, the entrapment of SDF in the resin-based fissure sealant did not potentiate the antibacterial effect of the fissure sealant or avoid the immediate development of dental caries. The development of more laboratorial research and, afterwards, long-term clinical data are necessary in order to verify if this association between these biomaterials is effective and can be considered for being used in oral health management. Also, other methodologies for associating cariostatic agents and sealant should be addressed.
Abstract: On 21st November, 2016, an outbreak of foodborne illness occurred after a buffet lunch served during a stakeholders’ consultation meeting held in Accra. An investigation was conducted to characterise the affected people, determine the etiologic food, the source of contamination and the etiologic agent and to implement appropriate public health measures to prevent future occurrences. A retrospective cohort study was conducted via telephone interviews, using a structured questionnaire developed from the buffet menu. A case was defined as any person suffering from symptoms of foodborne illness e.g. diarrhoea and/or abdominal cramps after eating food served during the stakeholder consultation meeting in Accra on 21st November, 2016. The exposure status of all the members of the cohort was assessed by taking the food history of each respondent during the telephone interview. The data obtained was analysed using Epi Info 7. An environmental risk assessment was conducted to ascertain the source of the food contamination. Risks of foodborne infection from the foods eaten were determined using attack rates and odds ratios. Data was obtained from 54 people who consumed food served during the stakeholders’ meeting. Out of this population, 44 people reported with symptoms of food poisoning representing 81.45% (overall attack rate). The peak incubation period was seven hours with a minimum and maximum incubation periods of four and 17 hours, respectively. The commonly reported symptoms were diarrhoea (97.73%, 43/44), vomiting (84.09%, 37/44) and abdominal cramps (75.00%, 33/44). From the incubation period, duration of illness and the symptoms, toxin-mediated food poisoning was suspected. The environmental risk assessment of the implicated catering facility indicated a lack of time/temperature control, inadequate knowledge on food safety among workers and sanitation issues. Limited number of food samples was received for microbiological analysis. Multivariate analysis indicated that illness was significantly associated with the consumption of the snacks served (OR 14.78, P < 0.001). No stool and blood or samples of etiologic food were available for organism isolation; however, the suspected etiologic agent was Staphylococcus aureus or Clostridium perfringens. The outbreak could probably be due to the consumption of unwholesome snack (tuna sandwich or chicken. The contamination and/or growth of the etiologic agent in the snack may be due to the breakdown in cleanliness, time/temperature control and good food handling practices. Training of food handlers in basic food hygiene and safety is recommended.
Abstract: Sulphur oxidizing bacteria (SOB) have marked their significant role in perspectives of maintaining healthy environment as researchers from all over the world tested and apply these in waste water treatment plants, bioleaching of heavy metals, deterioration of bridge structures, concrete and for bioremediation purposes, etc. Also, these SOB are well adapted in all kinds of environment ranging from normal soil, water habitats to extreme natural sources like geothermal areas, volcanic eruptions, black shale and acid rock drainage (ARD). SOB have been isolated from low pH environment of anthropogenic origin like acid mine drainage (AMD) and bioleaching heaps, hence these can work efficiently in different environmental conditions. Besides having many applications in field of environment science, they may be proven to be very beneficial in area of agriculture as sulphur is the fourth major macronutrients required for the growth of plants. More amount of sulphur is needed by pulses and oilseed crops with respect to the cereal grains. Due to continuous use of land for overproduction of more demanding sulphur utilizing crops and without application of sulphur fertilizers, its concentration is decreasing day by day, and thus, sulphur deficiency is becoming a great problem as it affects the crop productivity and quality. Sulphur is generally found in soils in many forms which are unavailable for plants (cannot be use by plants) like elemental sulphur, thiosulphate which can be taken up by bacteria and converted into simpler forms usable by plants by undergoing a series of transformations. So, keeping the importance of sulphur in view for various soil types, oilseed crops and role of microorganisms in making them available to plants, we made an effort to isolate, optimize, and characterize SOB. Three potential strains of bacteria were isolated, namely SSF7, SSA21, and SSS6, showing sulphate production of concentration, i.e. 2.268, 3.102, and 2.785 mM, respectively. Also, these were optimized for various culture conditions like carbon, nitrogen source, pH, temperature, and incubation time, and characterization was also done.
Abstract: Ancient Thai perfumed powder was used as a fragrance for clothing, food, and the body. Plant-based natural Thai perfume products are known as Pang-Rum. The objective of this study was to evaluate the stability of essential oils after six months of incubation. The chemical compositions were determined by gas chromatography-mass spectrometry (GC-MS), in terms of the qualitative composition of the isolated essential oil. The isolation of the essential oil of natural products by incubate sample for 5 min at 40 ºC is described. The volatile components were identified by percentage of total peak areas comparing their retention times of GC chromatograph with NIST mass spectral library. The results show no significant difference in the seven chromatograms of perfumed powder (Pang-Rum) both with binder and without binder. Further identification was done by GC-MS. Some components of Pang-Rum with/without binder were changed by temperature and time.
Abstract: Municipal Solid Waste (MSW), being a rich source of organic materials, can be used for agricultural applications as an important source of nutrients for soil and plants. This is also an alternative beneficial management practice for MSW generated in developing countries. In the present study, MSW treated soil samples from last four to six years at farmer’s field in Rohtak and Gurgaon states (Haryana, India) were collected. The samples were analyzed for all-important agricultural parameters and compared with the control untreated soil samples. The treated soil at farmer’s field showed increase in total N by 48 to 68%, P by 45.7 to 51.3%, and K by 60 to 67% compared to untreated soil samples. Application of sewage sludge at different sites led to increase in microbial biomass C by 60 to 68% compared to untreated soil. There was significant increase in total Cu, Cr, Ni, Fe, Pb, and Zn in all sewage sludge amended soil samples; however, concentration of all the metals were still below the current permitted (EU) limits. To study the adverse effect of heavy metals accumulation on various soil microbial activities, the sewage sludge samples (from wastewater treatment plant at Gurgaon) were artificially contaminated with heavy metal concentration above the EU limits. They were then applied to soil samples with different rates (0.5 to 4.0%) and incubated for 90 days under laboratory conditions. The samples were drawn at different intervals and analyzed for various parameters like pH, EC, total N, P, K, microbial biomass C, carbon mineralization, and diethylenetriaminepentaacetic acid (DTPA) exactable heavy metals. The results were compared to the uncontaminated sewage sludge. The increasing level of sewage sludge from 0.5 to 4% led to build of organic C and total N, P and K content at the early stages of incubation. But, organic C was decreased after 90 days because of decomposition of organic matter. Biomass production was significantly increased in both contaminated and uncontaminated sewage soil samples, but also led to slight increases in metal accumulation and their bioavailability in soil. The maximum metal concentrations were found in treatment with 4% of contaminated sewage sludge amendment.
Abstract: Aim of this study is to determine the effects of cellular insulin receptor stimulators on performance, plasma glucose, high density lipoprotein (HDL), low density lipoprotein (LDL), total cholesterol, triglyceride, triiodothyronine (T3) and thyroxine (T4) hormone levels, and incubation features in the breeding Japanese quails (Coturnix japonica). In the study, a total of 84 breeding quails was used, 6 weeks’ age, 24 are male and 60, female. Rations used in experiment are 2900 kcal/kg metabolic energy and 20% crude protein. Water pH is calibrated to 7.45. Ration and water were administered ad-libitum to the animals. As metformin source, metformin-HCl was used and as chrome resource, chromium picolinate was used. Trial groups were formed as control group (basal ration), metformin group (basal ration, added metformin at the level of feed of 20 mg/kg), and chromium picolinate (basal ration, added feed of 1500 ppb Cr) group. When regarded to the results of performance at the end of experiment, it is seen that live weight gain, feed consumption, egg weight, feed conversion ratio (Feed consumption/ egg weight), and egg production were affected at the significant level (p < 0.05). When the results are evaluated in terms of incubation features, hatchability and hatchability of fertile egg ratio were not affected from the treatments. Fertility ratio was significantly affected by metformin and chromium picolinate treatments and fertility rose at the significant level compared to control group (p < 0.05). According to results of experiment, plasma glucose level was not affected by metformin and chromium picolinate treatments. Plasma, total cholesterol, HDL, LDL, and triglyceride levels were significantly affected from insulin receptor stimulators added to ration (p < 0.05). Hormone level of Plasma T3 and T4 were also affected at the significant level from insulin receptor stimulators added to ration (p < 0.05).
Abstract: The present in vitro study was designed to reveal whether amygdalin (AMG) is able to cause changes to the motility, viability and mitochondrial activity of rabbit spermatozoa. New Zealand White rabbits (n = 10) aged four months were used in the study. Semen samples were collected from each animal and used for the in vitro incubation. The samples were divided into five equal parts and diluted with saline supplemented with 0, 0.5, 1, 2.5 and 5 mg/mL AMG. At times 0h, 3h and 5h spermatozoa motion parameters were assessed using the SpermVision™ computer-aided sperm analysis (CASA) system, cell viability was examined with the metabolic activity (MTT) assay, and the eosin-nigrosin staining technique was used to evaluate the viability of rabbit spermatozoa. All AMG concentrations exhibited stimulating effects on the spermatozoa activity, as shown by a significant preservation of the motility (P
Abstract: The environmental fate of organic contaminants in soils is influenced significantly by the pH, texture of soil, water content and also presence of organic matter. In this study, biodegradation of endosulfan isomers was studied in two different soils (Soil A and Soil B) that have contrasting properties in terms of their texture, pH, organic content, etc. Two Nocardia sp., which were isolated from soil, were used for degradation of endosulfan. Soils were contaminated with commercial endosulfan. Six sets were maintained from two different soils, contaminated with different endosulfan concentrations for degradation experiments. Inoculated and uninoculated mineral media with Nocardia isolates were added to the soils and mixed. Soils were incubated at a certain temperature (30 °C) during ten weeks. Residue endosulfan and its metabolites’ concentrations were determined weekly during the incubation period. The changes of the soil microorganisms were investigated weekly.
Abstract: The aim of this study was to estimate the digestibility of the fruit internal skin of different varieties of hazelnuts to propose hazelnut fruit skin as an alternative feed source as roughage in ruminant nutrition. In 2015, the fruit internal skins of three different varieties of round hazelnuts (RH), pointed hazelnuts (PH) and almond hazelnuts (AH) were obtained from hazelnut processing factory then their crude nutrients analysis were carried out. Organic matter digestibility (OMD) and metabolisable energy (ME) values of hazelnut fruit skins were estimated from gas measured by in vitro gas production method. Their antioxidant activities were determined by spectrophotometric method. Crude nutrient values of three different varieties were; organic matter (OM): 87.83, 87.81 and 87.78%), crude protein (CP): 5.97, 5.93 and 5.89%, neutral detergent fiber (NDF): 30.30, 30.29 and 30.29%, acid detergent fiber (ADF): 48.68, 48.67 and 48.66% and acid detergent lignin (ADL): 25.43, 25.43 and 25.39% respectively. OMD from 24 h incubation time of RH, PH and AH were 22.04, 22.46 and 22.74%; MEGP values were 3.69, 3.75 and 3.79 MJ/kg DM; and antioxidant activity values were 94.60, 94.54 and 94.52 IC 50 mg/mL respectively. The fruit internal skin of different varieties of hazelnuts may be considered as an alternative roughage for ruminant nutrition regarding to their crude and digestible nutritive values. Moreover, hazelnut fruit skin has a rich antioxidant content so it may be used as a feed additive for both ruminant and non-ruminant animals.
Abstract: Insufficient informal communication systems can lead to the first crisis (‘Crisis of Leadership’) for start-ups. Management Control Systems (MCS) are one way for high-technology start-ups to successfully overcome these problems. So far the literature has investigated the incubation of a start-up, but focused less on the post-incubation stage. This paper focuses on the use of MCS in post-incubation and, if failed start-ups agree, on how MCS are used. We conducted 14 semi-structured interviews for this purpose, to obtain our results. The overall conclusion is that the majority of the companies were closed down due to a combination of strategic, operative and financial reasons.
Abstract: Developing our knowledge of when pineapple roots
grow can lead to improved water, fertilizer applications, and more
precise culture management. This paper presents current
understanding of morphological traits in pineapple roots, highlighting
studies using incubation periods and various solid MS media treated
with different sucrose concentrations and pH, which directly assess in
vitro environmental factors. Rooting parameters had different optimal
sucrose concentrations and incubation periods. All shoots failed to
root in medium supplemented with sucrose at 5 g/L and no roots
formed within the first 45 days in medium enriched with sucrose at
10 g/L. After 75 days, all shoots rooted in medium enriched with 10
and 20 g/L sucrose. Moreover, MS medium supplied with 20 g/L
sucrose resulted in the longest and the highest number of roots with
27.3 mm and 4.7, respectively. Root function, such as capacity for P
and N uptake, declined rapidly with root length. As a result, the
longer the incubation period, the better the rooting responses would
be.
Abstract: Target of this study was the analysis of the impact of
crude glycerol on canine spermatozoa motility, morphology,
viability, and membrane integrity. Experiments were realized in vitro.
In the study, semen from 5 large dog breeds was used. They were
typical representatives of large breeds, coming from healthy rearing,
regularly vaccinated and integrated to the further breeding. Semen
collections were realized at the owners of animals and in the
veterinary clinic. Subsequently the experiments were realized at the
Department of Animal Physiology of the SUA in Nitra. The
spermatozoa motility was evaluated using CASA analyzer
(SpermVisionTM, Minitub, Germany) at the temperature 5 and 37°C
for 5 hours. In the study, 13 motility parameters were evaluated.
Generally, crude glycerol has generally negative effect on
spermatozoa motility. Morphological analysis was realized using
Hancock staining and the preparations were evaluated at
magnification 1000x using classification tables of morphologically
changed spermatozoa. Data clearly detected the highest number of
morphologically changed spermatozoa in the experimental groups
(know twisted tails, tail torso and tail coiling). For acrosome
alterations swelled acrosomes, removed acrosomes and acrosomes
with undulated membrane were detected. In this study also the effect
of crude glycerol on spermatozoa membrane integrity were analyzed.
The highest crude glycerol concentration significantly affects
spermatozoa integrity. Results of this study show that crude glycerol
has effect of spermatozoa motility, viability, and membrane integrity.
Detected changes are related to crude glycerol concentration,
temperature, as well as time of incubation.
Abstract: Past literature on business incubators distinguished incubators based on their mission statements. However, more and more mission statements become a slogan rather than a reality. It is therefore more appropriate to identify business incubators based on their real activities, rather than the missions they declared. With a sample of technology business incubators (TBIs) in China, we try to investigate business incubators’ real activities by examining the incubation efficiency along the following five dimensions, i.e., survival of new ventures, technology transfer, local economic growth, job creation, and profit generation. Furthermore, we identified six types of business incubators. The results indicate that generally Chinese TBIs have a greater preference for acquiring profits over other dimensions.
Abstract: In the immunologic sense, clinical infection is a state
of failure of the immune system to combat the pathogenic weapon of
the bacteria invading the host. A motile gram negative vibroid
organism associated with marked mono and poly nuclear cell
responses was traced during the examination of a clinical material
from an infected common carp Cyprinus carpio. On primary plate
culture, growth was shown to be pure, dense population of an
Aeromonas-like colony morphotype. The pure isolate was found to
be; Aerobic, facultatively anaerobic, non-halophilic, grew at 0C, and
37C, oxidase positive utilizes glucose through fermentative pathway,
resist 0/129 and novobiocin, produces alanine and lysine
decarboxylases but non-producing ornithine dehydrolases. Tests for
the in vitro determinants of pathogenicity has shown to be; Betahaemolytic
onto blood agar, gelatinase, casienase and amylase
producer. Three in vivo determinants of pathogenicity were tested as,
the lethal dose fifty, the pathogenesis and pathogenicity. It was
evident that 0.1 milliliter of the causal bacterial cell suspension of a
density 1 x 107 CFU/ml injected intramuscularly into an average of
100gms fish toke five days incubation period, then at the day six
morbidity and mortality were initiated. LD50 was recorded at the day
12 post-infection. Use of an LD50 doses to study the pathogenicity,
reveals mononuclear and polynuclear cell responses, on examining
the stained direct films of the clinical materials from the
experimentally infected fish. Re-isolation tests confirm that the reisolant
is same. The course of the infection in natural case was shown
manifestation of; skin ulceration, haemorrhage and descaling. On
evisceration, the internal organs were shown; congestion in the
intestines, spleen and, air sacs. The induced infection showed a
milder form of these manifestations. The grading of the virulence of
this organism was virulent causing chronic course of infections as
indicated from the pathogenesis and pathogenicity studies. Thus the
infectious bacteria were consistent with Aeromonas hydrophila, and
the infection was chronic.
Abstract: More than 3000 plants of notable phyto-therapeutic
value grow in South Africa; these include Cissampelos capensis,
commonly known in Afrikaans as dawidjie or dawidjiewortel. C.
capensis is the most significant and popular medicinal plant used by
the Khoisan as well as other rural groups in the Western region of
South Africa. Its rhizomes are traditionally used to treat male fertility
problems. Yet, no studies have investigated the effects of this plant or
its extracts on human spermatozoa. Therefore, this study aimed at
investigating the effects of C. capensis rhizome extract (CRE)
fractions on ejaculated human spermatozoa in vitro. Spermatozoa
from a total of 77 semen samples were washed with human tubular
fluid medium supplemented with bovine serum albumin (HTF-BSA)
and incubated for 2 hours with 20 μg/ml progesterone (P4) followed
by incubation with different concentrations (0, 0.05, 0.5, 5, 50, 200
μg/ml) of fractionated CRE (F1=0% MeOH, F2=30% MeOH,
F3=60% MeOH and F4=100% MeOH) for 1.5 hours at 37°C. A
sample without addition of CRE fractions served as control. Samples
were analyzed for sperm motility, reactive oxygen species (ROS),
DNA-fragmentation, acrosome reaction and capacitation. Results
showed that F1 resulted in significantly higher values for ROS,
capacitation and hyper-activation compared to F2, F3, and F4 with
P4-stimulated samples generally having higher values. No significant
effect was found for the other parameters. In conclusion, alkaloids
present in F1 of CRE appear to have triggered sperm intrinsic ROS
production leading to sperm capacitation and acrosome reaction
induced by P4.
Abstract: The potential neuroprotective effect of Phyllantus
nuriri against Fe2+ and sodium nitroprusside (SNP) induced oxidative
stress in mitochondria of rats brain was evaluated. Cellular viability
was assessed by MTT reduction, reactive oxygen species (ROS)
generation was measured using the probe 2,7-dichlorofluoresce
indiacetate (DCFH-DA). Glutathione content was measured using
dithionitrobenzoic acid (DTNB). Fe2+ (10μM) and SNP (5μM)
significantly decreased mitochondrial activity, assessed by MTT
reduction assay, in a dose-dependent manner, this occurred in parallel
with increased glutathione oxidation, ROS production and lipid
peroxidation end-products (thiobarbituric acid reactive substances,
TBARS). The co-incubation with methanolic extract of Phyllantus
nuriri (10-200 μg/ml) reduced the disruption of mitochondrial
activity, gluthathione oxidation, ROS production as well as the
increase in TBARS levels caused by both Fe2+ and SNP in a dose
dependent manner. HPLC analysis of the extract revealed the
presence of gallic acid (20.540.01), caffeic acid (7.930.02), rutin
(25.310.05), quercetin (31.280.03) and kaemferol (14.360.01).
This result suggests that these phytochemicals account for the
protective actions of P. niruri against Fe2+ and SNP -induced
oxidative stress. Our results show that P. nuriri consist important
bioactive molecules in the search for an improved therapy against the
deleterious effects of Fe2+, an intrinsic producer of reactive oxygen
species (ROS), that leads to neuronal oxidative stress and
neurodegeneration.