Search results for: zoospore inhibition assay

Authors: Andreina Giustiniani, Massimiliano Oliveri

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Rhythmic activity in different frequencies could subserve distinct functional roles during visual perception and visual mental imagery. In particular, alpha band activity is thought to play a role in active inhibition of both task-irrelevant regions and processing of non-relevant information. In the present blind placebo-controlled study we applied alpha transcranial alternating current stimulation (tACS) in the occipital cortex both during a basic visual perception and a visual working memory task. To understand if the role of alpha is more related to a general inhibition of distractors or to an inhibition of task-irrelevant regions, we added a non visual distraction to both the tasks.Sixteen adult volunteers performed both a simple perception and a working memory task during 10 Hz tACS. The electrodes were placed over the left and right occipital cortex, the current intensity was 1 mA peak-to-baseline. Sham stimulation was chosen as control condition and in order to elicit the skin sensation similar to the real stimulation, electrical stimulation was applied for short periods (30 s) at the beginning of the session and then turned off. The tasks were split in two sets, in one set distracters were included and in the other set, there were no distracters. Motor interference was added by changing the answer key after subjects completed the first set of trials.The results show that alpha tACS improves working memory only when no motor distracters are added, suggesting a role of alpha tACS in inhibiting non-relevant regions rather than in a general inhibition of distractors. Additionally, we found that alpha tACS does not affect accuracy and hit rates during the visual perception task. These results suggest that alpha activity in the occipital cortex plays a different role in perception and working memory and it could optimize performance in tasks in which attention is internally directed, as in this working memory paradigm, but only when there is not motor distraction. Moreover, alpha tACS improves working memory performance by means of inhibition of task-irrelevant regions while it does not affect perception.

Keywords: alpha activity, interference, perception, working memory

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Authors: H. Bouaziz, M. Sefi, J. de Lapuente, M. Borras, N. Zeghal

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Although arsenic trioxide has been the subject of toxicological research, in vitro cytotoxicity and genotoxicity studies using relevant cell models and uniform methodology are not well elucidated. Hence, the aim of the present study was to evaluate the cytotoxicity and genotoxicity induced by arsenic trioxide in human keratinocytes (HaCaT) using the MTT [3-(4, 5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide] and alkaline single cell gel electrophoresis (Comet) assays, respectively. Human keratinocytes were treated with different doses of arsenic trioxide for 4 h prior to cytogenetic assessment. Data obtained from the MTT assay indicated that arsenic trioxide significantly reduced the viability of HaCaT cells in a dose-dependent manner, showing a IC50 value of 34.18 ± 0.6 µM. Data generated from the comet assay also indicated a significant dose-dependent increase in DNA damage in HaCaT cells associated with arsenic trioxide exposure. We observed a significant increase in comet tail length and tail moment, showing an evidence of arsenic trioxide -induced genotoxic damage in HaCaT cells. This study confirms that the comet assay is a sensitive and effective method to detect DNA damage caused by arsenic.

Keywords: arsenic trioxide, cytotoxixity, genotoxicity, HaCaT

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Authors: Wenxiao Liu, Kun Zhang, Yongqing Li

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Bovine herpesvirus 1, a member of the genus Variellovirus of the subfamily Alphaherpesvirinae, has caused severe economic cost to the bovine industry. In this study, BoHV-1 glycerol protein gD was expressed in insect cells, and the purified gD was immunized in the Balb/C mice to generate monoclonal antibodies. Based on hybridoma cell fusion techniques, 20 monoclonal antibodies against Bovine herpesvirus 1 have been obtained. Further, mAb 3F8 with neutralizing activity and gD were applied to develop a blocking enzyme-linked immunosorbent assay (Elisa) for detecting neutralizing antibodies against BoHV-1, which shows a significant correlation between the blocking Elisa and VNT. The sensitivity and specificity of the test were estimated to be 94.59% and 93.42%, respectively. Furthermore, antibody pairing tests revealed that mAb 1B6 conjugated to fluorescence microspheres was used as the capture antibody, and mAb 3F9 was used as the detectable antibody to establish the immunochromatographic assay (ICS). The ICS was conducted to detect BoHV-1 in bovine samples with high sensitivity, specificity, and good stability. Clinical sample testing revealed that the results of ICS and real-time PCR have a coincidence rate of 95.42%. Our research confirmed that the ICS is a rapid and reliable method for the diagnosis of BoHV-1. In conclusion, our results lay a solid foundation for the prevention and control of BoHV-1 infection.

Keywords: bovine disease, BoHV-1, ELISA, ICS assay

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Authors: Kai-Fen Tu, Jenn-Wen Huang, Yao-Tung Lin

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Cucumber is a major economic crop in the world. The global production of cucumber in 2017 was more than 71 million tonnes. Nonetheless, downy mildew, caused by Pseudoperonospora cubensis, is a devastating and common disease on cucumber in around 80 countries and causes severe economic losses. The long-term usage of fungicide also leads to the occurrence of fungicide resistance and decreases host resistance. In this study, six types of oil (neem oil, moringa oil, soybean oil, cinnamon oil, clove oil, and camellia oil) were selected to synthesize micro/nano-sized emulsions, and the disease control efficacy of micro/nano-sized emulsions were evaluated. Moreover, oil concentrations (0.125% - 1%) and droplet size of emulsion were studied. Results showed cinnamon-type emulsion had the best efficacy among these oils. The disease control efficacy of these emulsions increased as the oil concentration increased. Both disease incidence and disease severity were measured by detached leaf and pot experiment, respectively. For the droplet size effect, results showed that the 114 nm of droplet size synthesized by 0.25% cinnamon oil emulsion had the lowest disease incidence (6.67%) and lowest disease severity (33.33%). The release of zoospore was inhibited (5.33%), and the sporangia germination was damaged. These results suggest that cinnamon oil emulsion will be a valuable and environmentally friendly alternative to control cucumber downy mildew. The economic loss caused by plant disease could also be reduced.

Keywords: downy mildew, emulsion, oil droplet size, plant protectant

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Authors: H. Minor-Pérez, A. M. Mota-Silva, S. Ortiz-Barrios

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Substances of vegetable origin with antioxidant capacity have a high potential for application on the conservation of some foods, can prevent or reduce for example oxidation of lipids. However a food is a complex system whose wide variety of components wich can reduce or eliminate this antioxidant capacity. The antioxidant activity can be determined with the ABTS technique. The radical ABTS+ is generated from the acid 2, 2´ - Azino-bis (3-ethylbenzothiazoline-6-sulfonic acid) (ABTS). This radical is a composite color bluish-green, stable and with a spectrum of absorption into the UV-visible. The addition of antioxidants causes discoloration, value that can be reported as a percentage of inhibition of the cation radical ABTS+. The objective of this study was evaluated the effect of the combination of the pH and the essential oil of cinnamon (EOC) on inhibition of the radical ABTS+, using statistical design of central point (Design Expert) to obtain mathematical models that describe this phenomenon. Were evaluated 17 treatments with combinations of pH 5, 6 and 7 (citrate-phosphate buffer) and the concentration of essential oil of cinnamon (C): 0 µg/mL, 100 µg/mL and 200 µg/mL. The samples were analyzed using the ABTS technique. The reagent was dissolved in methanol 80% to standardized the absorbance to 0.7 +/- 0.1 at 754 nm. Then samples were mixed with reagent standardized ABTS and after 1 min and 7 min absorbance was read for each treatment at 754 nm. Was used a curve pattern with vitamin C and reported the values as inhibition (%) of radical ABTS+. The statistical analysis shows the experimental results were adjusted to a quadratic model, to the times of 1 min and 7 min. This model describes the influence of the factors investigated independently: pH and cinnamon essential oil (µg/mL) and the effect of the interaction between pH*C, as well as the square of the pH2 and C2. The model obtained was Y = 10.33684 - 3.98118*pH + 1.17031*C + 0.62745*pH2 - 3.26675*10-3*C2 - 0.013112*pH*C, where Y is the response variable. The coefficient of determination was 0.9949 for 1 min. The equation was obtained at 7 min and = - 10.89710 + 1.52341*pH + 1.32892*C + 0.47953*pH2 - 3.56605*10- *C2 - 0.034687*pH*C. The coefficient of determination was 0.9970. This means that only 1% of the total variation is not explained by the developed models. At 100 µg/mL of EOC was obtained an inhibition percentage of 80%, 84% and 97% for the pH values of 5,6 and 7 respectively, while a value of 200 µg/mL the inhibition (%) was very similar for the treatments. In these values of pH was obtained an inhibition close 97%. In conclusion the pH does not have a significant effect on the antioxidant capacity, while the concentration of EOC was decisive for the antioxidant capacity. The authors acknowledge the funding provided by the CONACYT for the project 131998.

Keywords: antioxidant activity, ABTS technique, essential oil of cinnamon, mathematical models

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Authors: Neha Singh, Anuj Ranjan, Tanu Jindal

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Over the last decade, the exponential growth of mobile communication has been accompanied by a parallel increase in density of electromagnetic fields (EMF). The continued expansion of mobile phone usage raises important questions as EMF, especially radio frequency (RF), have long been suspected of having biological effects. In the present experiments, we studied the effects of RF-EMF on cell death (apoptosis) and DNA damage of a well- tested biological model, Drosophila melanogaster exposed to 2400 MHz frequency for different time duration i.e. 2 hrs, 4 hrs, 6 hrs,8 hrs, 10 hrs, and 12 hrs each day for five continuous days in ambient temperature and humidity conditions inside an exposure chamber. The flies were grouped into control, sham-exposed, and exposed with 100 flies in each group. In this study, well-known techniques like Comet Assay and TUNEL (Terminal deoxynucleotide transferase dUTP Nick End Labeling) Assay were used to detect DNA damage and for apoptosis studies, respectively. Experiments results showed DNA damage in the brain cells of Drosophila which increases as the duration of exposure increases when observed under the observed when we compared results of control, sham-exposed, and exposed group which indicates that EMF radiation-induced stress in the organism that leads to DNA damage and cell death. The process of apoptosis and mutation follows similar pathway for all eukaryotic cells; therefore, studying apoptosis and genotoxicity in Drosophila makes similar relevance for human beings as well.

Keywords: cell death, apoptosis, Comet Assay, DNA damage, Drosophila, electromagnetic fields, EMF, radio frequency, RF, TUNEL assay

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Authors: Laura Boschis, Elena D. Ozzello, Enzo Mastromatteo

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Point of care diagnostic finds growing interest in medicine and agri-food because of faster intervention and prevention. EliChip is a microfluidic platform to perform Point of Care immunoenzymatic assay based on ready-to-use kits and a portable instrument to manage fluidics and read reliable quantitative results. Thanks to miniaturization, analyses are faster and more sensible than conventional ELISA. EliChip is one of the crucial assets of the Europen-founded Flamingo project for in-line measuring inflammatory markers.

Keywords: lab on chip, point of care, immunoenzymatic analysis, synovial arthritis

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Authors: Nabila N. El-Maraghy, Amany Essa, Yousra Abdel–Mottaleb, Nada Ismail

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The use of combination of chemotherapy and natural products to influence the cell death with low doses of chemotherapeutic agents and few side effects has recently emerged as a new method of cancer therapy. Aim: Evaluation the modulatory effect of Thymoquinone on HepG2 cells treated with Sorafenib. Methods: Hepatocellular Carcinoma HepG2 cell line was treated with Sorafenib and TQ individually and in combination. The effect of these treatments on cell viability (MTT assay), apoptosis (Expression of Caspase-3) and oxidative markers (GSH content and extent of lipid peroxidation) was determined. Results: When compared the effect of both agents alone and the combination of the IC50 of Sorafenib and the IC50 TQ, the combination resulted in reduction of cell inhibition and apoptosis and antagonize their actions on GSH content and extent of lipid peroxidation which are increased. This study showed potent anti-tumor activity of both TQ and Sorafenib separately on HepG2 but upon combination surprisingly they interacted and give antagonistic effect. Conclusion: Co-treatment resulted in antagonistic interaction between Sorafenib and Thymoquinone.

Keywords: antagonism, hepatocellular carcinoma, sorafenib, thymoquinone

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Authors: Nurjannatul Naim Kamaruddin, Tengku Sifziuzl Tengku Muhammad, Aina Farahiyah Abdul Manan, Habsah Mohamad

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Atherosclerosis which leads to cardiovascular diseases such as myocardial infarction, unstable angina (ischemic heart pain), sudden cardiac death and stroke is the principal cause of death worldwide. It has been a very critical issue as current common drug treatment, statin therapy has left bad side effects like rhabdomyolysis, atrial fibrillation, liver disease, abdominal and chest pain. Interestingly, the discoveries of proprotein convertase subtilisin-kexin type 9 have paved a new way in the treatment of atherosclerosis. This serine protease is believed to involve in the regulation of LDL- uptake by LDL-receptor. Therefore, this study was conducted to evaluate the potential of Acanthaster plancii fractions to reduce the transcriptional activity of the PCSK9 promoter. In this study, the marine organism which is Acanthaster plancii has been used as the source for marine compounds in inhibiting PCSK9. The cytotoxicity activity of ten fractions from the methanol extracts of Acanthaster plancii was investigated on HepG2 cell lines using MTS assay and dual glo luciferase assay was carried out later to analyses the effects of the samples in reducing the transcriptional activity of the PCSK9 promoter. Both assays used fractions with five different concentrations, 3.13µg/mL, 6.25µg/mL, 12.5µg/mL, 25µg/mL, and 50µg/mL. MTS assay indicated that the fractions are non-cytotoxic towards HepG2 cell lines as their IC50 value is greater than 30µg/mL. Whilst, for the dual glo luciferase assay, among all the fractions, Enhance Fraction 2 (EF2) showed the best potential in reducing the transcriptional activity of the PCSK9 promoter. The results indicated that this EF2 gave the lowest PCSK9 promoter expression at low concentration which is 0.2 fold change at 6.25µg/mL. This finding suggested that further analysis should be done to validate the potential of Acanthaster plancii as the source of anti-atherosclerotic agent.

Keywords: Acanthaster plancii, atherosclerosis, luciferase assay, PCSK9

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Authors: Faraj M. Elkhatri, Hana Ellafi

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In recent years, there has been a growing recognition of the potential of marine-based functional foods and combination therapies in promoting a healthy lifestyle and exploring their effectiveness in preventing or treating diseases. The combination of marine bioactive compounds or extracts offers synergistic or enhancement effects through various mechanisms, including multi-target actions, improved bioavailability, enhanced bioactivity, and mitigation of potential adverse effects. Both the green-lipped mussel (GLM) and fucoidan derived from brown seaweed are rich in bioactivities. These two, mussel and fucoidan, have not been previously formulated together. This study aims to combine GLM oil from Perna canaliculus with low molecular weight fucoidan (LMWF) extracted from Undaria pinnatifida to investigate the unique mixture’s anti-inflammatory and antioxidant properties. The cytotoxicity of individual compounds and combinations was assessed using the MTT assay in (THP-1 and RAW264.7) cell lines. The anti-inflammatory activity of mussel-fucoidan was evaluated by treating LPS-stimulated human monocyte and macrophage (THP1-1) cells. Subsequently, the inflammatory cytokines released into the supernatant of these cell lines were quantified via ELISA. Antioxidant activity was determined by using the free radical scavenging assay (DPPH). DPPH assay demonstrated that the radical scavenging activity of the combinations, particularly at concentrations exceeding 1 mg/ml, showed a significantly higher percentage of inhibition when compared to the individual component. This suggests an enhancement effect when the two compounds are combined, leading to increased antioxidant activity. In terms of immunomodulatory activity, the individual compounds exhibited distinct behaviors. GLM oil displayed a higher ability to suppress the cytokine TNF- compared to LMWF. Interestingly, the LMWF fraction, when used individually, did not demonstrate TNF- suppression. However, when combined with GLM, the TNF- suppression (anti-inflammatory) activity of the combination was better than GLM or LWMF alone. This observation underscores the potential for enhancement interactions between the two components in terms of anti-inflammatory properties. This study revealed that each individual compound, LMWF, and GLM, possesses unique and notable bioactivity. The combination of these two individual compounds results in an enhancement effect, where the bioactivity of each is enhanced, creating a superior combination. This suggests that the combination of LMWF and GLM has the potential to offer a more potent and multifaceted therapeutic effect, particularly in the context of antioxidant and anti-inflammatory activities. These findings hold promise for the development of novel therapeutic interventions or supplements that harness the enhancement effects.

Keywords: formation damage, porosity loses, pore throat, quartz cement

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Authors: Brizita Djordjevic, Bojana Vidovic, Milica Zrnic, Uros Cakar, Ivan Stankovic, Davor Korcok, Sladjana Sobajic

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Histamine is a biogenic amine, which is formed by enzymatic decarboxylation from the amino acid histidine. It can be found in foods such as fish and fish products, meat and fermented meat products, cheese, wine and beer. The presence of histamine in these foods can indicate microbiological spoilage or poor manufacturing processes. The consumption of food containing large amounts of histamine can have toxicological consequences. In 62 food products (31 canned fish products, 19 wines and 12 cheeses) from the market of Serbia the content of histamine was determined using enzyme-linked immunosorbent assay (ELISA) test kit according to the manufacturer's instructions (Immunolab GmbH, Kassel, Germany). The detection limits of this assay were 20 µg/kg for fish and cheese and 4 µg/L for wine. The concentration of histamine varied between 0.16-207 mg/kg in canned fish products, 0.03-1.47 mg/kg in cheeses and 0.01- 0.18 mg/L in wines. In all analyzed canned fish products the results obtained for the histamine were below the limits set by European and national legislation, so they can be considered acceptable and safe for the health consumers. The levels of histamine in analyzed cheeses and wines were very low and did not pose safety concerns.

Keywords: cheese, enzyme-linked immunosorbent assay, histamine, fish products, wine

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Authors: Emma O’ Keeffe, Helen Hughes, Peter McLoughlin, Shiau Pin Tan, Nick McCarthy

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Biosecurity is emerging as one of the most important issues facing the agricultural and forestry community. This is as a result of increased invasion from new pests and diseases with the main protocol for dealing with these species being the use of synthetic pesticides. However, these chemicals have been shown to exhibit negative effects on the environment. Seaweeds represent a vast untapped resource of bio-molecules with a broad range of biological activities including pesticidal. This project investigated both the antifungal and antibacterial activity of seaweed species against two problematic root rot fungi, Armillaria mellea and Heterobasidion annosum and ten quarantine bacterial plant pathogens including Xanthomonas arboricola, Xanthomonas fragariae, and Erwinia amylovora. Four seaweed species were harvested from the South-East coast of Ireland including brown, red and green varieties. The powdered seaweeds were extracted using four different solvents by liquid extraction. The poisoned food technique was employed to establish the antifungal efficacy, and the standard disc diffusion assay was used to assess the antibacterial properties of the seaweed extracts. It was found that extracts of the green seaweed exhibited antifungal activity against H. annosum, with approximately 50% inhibition compared to the negative control. The protectant activities of the active extracts were evaluated on disks of Picea sitchensis, a plant species sensitive to infection from H. annosum and compared to the standard chemical control product urea. The crude extracts exhibited very similar activity to the 10% and 20% w/v concentrations of urea, demonstrating the ability of seaweed extracts to compete with commercially available products. Antibacterial activity was exhibited by a number of seaweed extracts with the red seaweed illustrating the strongest activity, with a zone of inhibition of 15.83 ± 0.41 mm exhibited against X. arboricola whilst the positive control (10 μg/disk of chloramphenicol) had a zone of 26.5 ± 0.71 mm. These results highlight the potential application of seaweed extracts in the forestry and agricultural industries for use as biopesticides. Further work is now required to identify the bioactive molecules that are responsible for this antifungal and antibacterial activity in the seaweed extracts, including toxicity studies to ensure the extracts are non-toxic to plants and humans.

Keywords: antibacterial, antifungal, biopesticides, seaweeds

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Authors: Fatiha Benghanem, Mokhtar Berarma, Saida Keraghel, Ali Ourari

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Corrosion is the result of the reaction between a material and its environment. Steel in concrete is protected from corrosion by a passive film promoted by concrete alkalinity. For the initiation of corrosion, this protective film must be destroyed and this can be mainly done in two ways: by the attack of chlorides on the steel or by carbonation of the cover concrete due the reaction with carbon dioxide, which causes reduction in the alkalinity of concrete. The literature reports several ways to decrease or to prevent reinforcement corrosion. Among them, the use of corrosion inhibitors has been an envisaged solution. Two approaches are generally used to evaluate the efficiency of inhibitors for concrete application; one uses simulated pore solution testing , and the other uses actual concrete or mortar specimens. Both methods are some times used in conjunction. The aim of this study is to investigate the use of 2,3-diaminopyridine as a corrosion inhibitors of steel in alkaline media which simulate the electrolyte in the concrete pores. The effectiveness of this compound as corrosion inhibitor was investigated by measuring the corrosion potentials, the polarization curves and the corrosion current densities of steel with and without chlorides. The study of corrosion inhibition by this compound led to the conclusion that he has low rates of inhibition in the absence of aggressive ions and high rates in their presence. This type of organic compounds are promoting for the protection of armatures in concrete.

Keywords: corrosion, inhibitors, mild steel, conjunction

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Authors: Shishen Xie, Yingda L. Xie

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Tuberculosis (TB) is a potentially serious infectious disease that remains a health concern. The Interferon Gamma Release Assay (IGRA) is a blood test to find out if an individual is tuberculous positive or negative. This study applies statistical analysis to the clinical data of interferon-gamma levels of seventy-three subjects who diagnosed pulmonary TB in an anti-tuberculous treatment. Data analysis is performed to determine if there is a significant decline in interferon-gamma levels for the subjects during a period of six months, and to infer if the anti-tuberculous treatment is effective.

Keywords: data analysis, interferon gamma release assay, statistical methods, tuberculosis infection

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Authors: Benyapa Suksuwan

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Introduction: Oxidative stress is identified as the root cause of the development and progression of several diseases as the disproportion of free radicals in the body leads to tissue or cell damage. Polyphenols are the most common antioxidant found in plants and are efficient in capturing oxidative free radicals. Aim of the Study: This study focused on the antioxidant activity of polyphenols extracted from Phyllanthus Emblica L. as oxidative stress plays a vital role in developing and progressing many diseases, including cardiovascular diseases and cancer. Materials and Methods: The plant was extracted using a mixture solvent (ethyl alcohol: water in ratio 8:2). The total phenolic content of P. Emblica extract was determined using the Folin-Cioucalteu method and calculated as gallic acid equivalents (GAE) and various antioxidant assays DPPH and ABTS radical scavenging capacity assays. Results and Discussion: The findings exhibited a strong correlation between antioxidant activity and the total phenol contents. In addition, the IC₅₀ of P. Emblica extract via DPPH and ABTS assays were 68.10 μg/mL ± 0.455, and 49.24 μg/mL ± 0.716, respectively. Furthermore, P. Emblica extract showed antioxidant activities in a concentration-dependent manner. Vitamin C was used as a positive control in the DPPH assay, while Trolox was used as a positive control in the ABTS assay. Conclusions: In conclusion, P. Emblica extract consisted of a high amount of total phenolic content, which possesses potent antioxidant activity. However, further antioxidant activity assays using human cell lines such as SOD, ROS, and RNS scavenging assays and in vitro antioxidant experiments should be performed in order.

Keywords: antioxidant, ABTS scavenging, DPPH scavenging assay, total phenol contents assay, Phyllanthus Emblica L

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Authors: Boutemak Khalida, Dahmani Siham

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Garden cress (Lepidium Sativum L.) belonging to the family Brassicaceae, is edible growing annual herb. Its various parts (roots, leaves and seeds) have been used to treat various human ailments. Its seed extracts have been screened for various biological activities like hypotensive, antimicrobial, bronchodilator, hypoglycaemic and antianemic. The aim of the present study is to optimize the process parameters (cellulase concentration and incubation time) of enzymatic pre-treatment of the garden cress seeds and to evaluate the effect of cellulase pre-treatment of the crushed seeds on the oil yield, physico-chemical properties and antibacterial activity and comparing to non-enzymatic method. The optimum parameters of cellulase pre-treatment were as follows: cellulase of 0,1% w/w and incubation time of 2h. After enzymatic pre-treatment, the oil was extracted by n-hexane for 1.5 h, the oil yield was 4,01% for cellulase pre-treatment as against 10,99% in the control sample. The decrease in yield might be caused a result of mucilage. Garden cress seeds are covered with a layer of mucilage which gels on contact with water. At the same time, the antibacterial activity was carried out using agar diffusion method against 4 food-borne pathogens (Escherichia coli, Salmonella typhi,Staphylococcus aureus, Bacillus subtilis). The results showed that bacterial strains are very sensitive to the oil with cellulase pre-treatment. Staphylococcus aureus is extremely sensitive with the largest zone of inhibition (40 mm), Escherichia coli and salmonella typhi had a very sensitive to the oil with a zone of inhibition (26 mm). Bacillus subtilizes is averagely sensitive which gave an inhibition of 16 mm. But it does not exhibit sensivity to the oil without enzymatic pre-treatment with a zone inhibition (< 8 mm). Enzymatic pre-treatment could be useful for antimicrobial activity of the oil, and hold a good potential for use in food and pharmaceutical industries.

Keywords: Lepidium sativum L., cellulase, enzymatic pretreatment, antibacterial activity.

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Authors: Mohammed Falalu Hamza

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A phytochemical investigation conducted on the leaves extract of Cryptocarya latifolia (Lauraceae) revealed the presence of two major essential oils; Nerolidol (1) and Copaene (2) with the aid of gas chromatography-mass spectrometry (GC-MS). The compounds exhibited good anti-oxidant capacity using 2,2-diphenyl-1-picryl-hydrazyl (DPPH) radical scavenging assay. The result shows that the anti-oxidant capacity of the compounds is dependent on concentration similar to the standard (ascorbic acid). This study shows that the leaves extract of C. latifolia is a good source of important natural antioxidants.

Keywords: broad-leaved quince, phytochemical, anti-oxidant, essential oils

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Authors: Abdolreza Esmaeilzadeh, Maryam Erfanmanesh, Sousan Ghasemi, Farzaneh Mohammadi

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Background: Hepatitis C Virus (HCV), a public health problem, is an enveloped, single-stranded RNA virus and a member of the Hepacivirus genus of the Flaviviridae family. Liver cancer, cirrhosis, and end-stage liver are the outcomes of chronic infection with HCV. HCV isolates show significant heterogeneity in genetics around the world. Therefore, determining HCV genotypes is a vital step in determining prognosis and planning therapeutic strategies. Materials and Methods: Serum samples of 136 patients were collected and analyzed for anti-HCV antibodies using ELISA (The enzyme-linked immunosorbent assay) method. Then, positive samples were exposed to RT-PCR, which was performed under standard condition. Afterwards, they investigated for genotyping using allele-specific PCR (AS-PCR), and HCV genotype 2.0 line probe assay (LiPA). Results: Samples indicated 216 bp bands on 2% agarose gel. Analyses of the results demonstrated that the most dominant subtype was 3a with frequency of 38.26% in Zanjan Province followed by subtypes of 1b, 1a, 2, and 4 with frequencies of 25.73%, 22.05%, 5.14%, and 4.41%, respectively. The frequency of unknown HCV genotypes was 4.41%. Conclusions: According to the results, it was found that HCV high prevalent genotype in Zanjan is subtype 3a. Analysis of the results provides identification of certain HCV genotypes, and these valuable findings could affect the type and duration of the treatment.

Keywords: anti-HCV antibody, Hepatitis C Virus (HCV), genotype, RT-PCR, AS-PCR

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Authors: M. N. Hanina, M. Hairul Shahril, I. Ismatul Nurul Asyikin, A. K. Abdul Jalil, M. R. Salina, M. R. Maryam, M. Rosfarizan

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The extracellular proteins secreted by bacteria may be increased in stressful surroundings, such as in the presence of antibiotics. It appears that many antibiotics, when used at low concentrations, have in common the ability to activate or repress gene transcription, which is distinct from their inhibitory effect. There have been comparatively few studies on the potential of antibiotics as a specific chemical signal that can trigger a variety of biological functions. Therefore, this study was carried out to determine the effect of Streptomycin Sulfate in regulating extracellular proteins secreted by Bacillus subtilis ATCC21332. Results of Microdilution assay showed that the Minimum Inhibition Concentration (MIC) of Streptomycin Sulfate on B. subtilis ATCC21332 was 2.5 mg/ml. The bacteria cells were then exposed to Streptomycin Sulfate at concentration of 0.01 MIC before being further incubated for 48h to 72 h. The extracellular proteins secreted were then isolated and analyzed by sodium dodecyl sulfate polyacrylamide gel electrophoresis (SDS-PAGE). Proteins profile revealed that three additional bands with approximate sizes of 30 kDa, 22 kDa and 23 kDa were appeared for the treated bacteria with Streptomycin Sulfate. Thus, B. subtilis ATCC21332 in stressful condition with the presence of Streptomycin Sulfate at low concentration could induce the extracellular proteins secretion.

Keywords: Bacillus subtilis ATCC21332, streptomycin sulfate, extracellular proteins, antibiotics

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Authors: N. A. Kazemi Sefat, M. M. Mohammadi, J. Hadjati, S. Talebi, M. Ajami, H. Daneshvar

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Colorectal cancer metastases result in a significant number of cancer related deaths. Histone deacetylase (HDAC) inhibitors induce growth arrest and apoptosis in a variety of human cancer cells. Sodium butyrate (SB) is a short chain fatty acid, belongs to HDAC inhibitors which is released in the colonic lumen as a consequence of fiber fermentation. In this study, we are about to assess the effect of sodium butyrate on HCT116 human colorectal carcinoma cell line. The viability of cells was measured by microscopic morphologic study and MTT assay. After 48 hours, treatments more than 10 mM lead to cell injury in HCT116 by increasing cell granulation and decreasing cell adhesion (p>0.05). After 72 hours, treatments at 10 mM and more lead to significant cell injury (p<0.05). Our results may suggest that the gene expression which is contributed in cell proliferation and apoptosis has been changed under pressure of HDAC inhibition.

Keywords: colorectal cancer, sodium butyrate, cytotoxicity, MTT

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Authors: Ana M. T. Mata, Alexiane Ligneul

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Textile effluents are usually heavily loaded with organic carbon and color compounds, the latter being azo dyes in an estimated 70% of the case effluent posing a major challenge in environmental protection. In this study, the ecotoxicity of simulated textile effluent after biological treatment with anaerobic and aerobic phase (aerobic granular sludge, AGS) and after advanced oxidation processes (AOP) namely ozonation and UV irradiation as post-treatment, were tested to evaluate the fitness of this treatments for ecotoxicity abatement. AGS treatment achieved an 80% removal in both COD and color. AOP was applied with the intention to mineralize the metabolites resulting from biodecolorization of the azo dye Acid Red 14, especially the stable aromatic amine (4-amino-1-naphthalenesulfonic acid, 4A1NS). The ecotoxicity evaluation was based on growth inhibition of the algae Pseudokirchneriella subcapitata following OECD TG 201 except regarding the medium, MBL medium was used instead. Five replicate control cultures and samples were performed with an average STD of 2.7% regarding specific algae growth rate determination. It was found that untreated textile effluent holds an inhibition of specific growth rate of 82%. AGS treatment by itself is able to lower ecotoxicity to 53%. This is probably due to the high color removal of the treatment. AOP post-treatment with Ozone and UV irradiation improves the ecotoxicity abatment to 49 and 43% inhibition respectively, less significantly than previously thought. Since over 85% of 4A1NS was removed by either of the AOP (followed by HPLC), an individual ecotoxicity test of 4A1NS was performed showing that 4A1NS does not inhibit algae growth (0% inhibition). It was concluded that AGS treatment is able by itself to achieve a significant ecotoxicity abatement of textile effluent. The cost-benefit of AOP as a post-treatment have to be better accessed since their application resulted in an improvement of only 10% regarding ecotoxicity effluent removal. It was also found that the 4A1NS amine had no apparent effect on ecotoxicity. Further studies will be conducted to study where ecotoxicity is coming from after AGS biological treatment and how to eliminate it.

Keywords: textile wastewate, ecotoxicity, aerobic granular sludge, AOP

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Authors: Suharni Mohamad Suharni Mohamad, Nurul Izzati Hamzan Nurul Izzati Hamzan, Norhayu Abdul Rahman Norhayu Abdul Rahman, Siti Suraiya Md Noor Siti Suraiya Md Noor

Abstract:

Human papillomavirus (HPV) is an important risk factor for development of oral cancer. HPV16 is the most common type found in HPV-positive squamous cell carcinoma. In the present study, we established a real-time loop-mediated isothermal amplification (real-time LAMP) for detection of HPV16. A set of six primers was specially designed to recognize eight distinct sequences of HPV16-E6. Detection and quantification was achieved by real-time monitoring using a real-time turbidimeter based on threshold time required for turbidity in the LAMP reaction. LAMP reagents (MgSO4, dNTPs, Bst polymerase concentrations) and various incubation times and temperatures were optimized. The sensitivity was determined using 10-fold serial dilutions of HPV16 standard strain. The specificity of was evaluated using other HPV genotypes. The optimized method was established with specifically designed primers by real-time detection in approximately 30 min at 65°C. The limit of detection of HPV16 using the LAMP assay was 10 pg/ml that could be detected in 30 min. The LAMP assay was 10 times more sensitive than the conventional PCR in detecting HPV16. No cross-reactivity with other HPV genotypes was observed. This quantitative real-time LAMP assay may improve diagnostic potential for the detection and quantification of HPV16 in clinical samples and epidemiological studies due to its rapidity, simplicity, high sensitivity and specificity. This assay will be further evaluated with HPV DNAs of saliva from patients with oral squamous cell carcinoma. Acknowledgement: This study was financially supported by the ScienceFund Grant, Ministry of Science, Technology and Innovation (305/PPSG/6113219).

Keywords: Oral Squamous Cell Carcinoma (OSCC), Human Papillomavirus 16 (HPV16), Loop-Mediated Isothermal Amplification (LAMP), rapid detection

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Authors: Yogesh Dalvi, Ruby Varghese, Nibu Varghese, C. K. Krishnan Nair

Abstract:

Introduction: The Genus Phellinus, locally known as Phansomba is a well-known traditional folk medicine. Especially, in Western Ghats of India, many tribes use several species of Phellinus for various ailments related to teeth, throat, tongue, stomach and even wound healing. It is one of the few mushrooms which play a pivotal role in Ayurvedic Dravyaguna. Aim: The present study focuses on to investigate phytochemical analysis, antioxidant, and antitumor (in vitro and in vivo) potential of Phellinus robinae from South India, Kerala Material and Methods: The present study explores the following: 1. Phellinus samples were collected from Ranni, Pathanamthitta district of Kerala state, India from Artocarpus heterophyllus Lam. and species were identified using rDNA region. 2. The fruiting body was shadow dried, powdered and extracted with 50% alcohol using water bath at 60°C which was further condensed by rotary evaporator and lyophilized at minus 40°C temperature. 3. Secondary metabolites were analyzed by using various phytochemical screening assay (Hager’s Test, Wagner’s Test, Sodium hydroxide Test, Lead acetate Test, Ferric chloride Test, Folin-ciocalteu Test, Foaming Test, Benedict’s test, Fehling’s Test and Lowry’s Test). 4. Antioxidant and free radical scavenging activity were analyzed by DPPH, FRAP and Iron chelating assay. 5. The antitumor potential of Water alcohol extract of Phellinus (PAWE) is evaluated through In vitro condition by Trypan blue dye exclusion method in DLA cell line and In vivo by murine model. Result and Discussion: Preliminary phytochemical screening by various biochemical tests revealed presence of a variety of active secondary molecules like alkaloids, flavanoids, saponins, carbohydrate, protein and phenol. In DPPH and FRAP assay PAWE showed significantly higher antioxidant activity as compared to standard Ascorbic acid. While, in Iron chelating assay, PAWE exhibits similar antioxidant activity that of Butylated Hydroxytoluene (BHT) as standard. Further, in the in vitro study, PAWE showed significant inhibition on DLA cell proliferation in dose dependent manner and showed no toxicity on mice splenocytes, when compared to standard chemotherapy drug doxorubicin. In vivo study, oral administration of PAWE showed dose dependent tumor regression in mice and also raised the immunogenicity by restoring levels of antioxidant enzymes in liver and kidney tissue. In both in vitro and in vivo gene expression studies PAWE up-regulates pro-apoptotic genes (Bax, Caspases 3, 8 and 9) and down- regulates anti-apoptotic genes (Bcl2). PAWE also down regulates inflammatory gene (Cox-2) and angiogenic gene (VEGF). Conclusion: Preliminary phytochemical screening revealed that PAWE contains various secondary metabolites which contribute to its antioxidant and free radical scavenging property as evaluated by DPPH, FRAP and Iron chelating assay. PAWE exhibits anti-proliferative activity by the induction of apoptosis through a signaling cascade of death receptor-mediated extrinsic (Caspase8 and Tnf-α), as well as mitochondria-mediated intrinsic (caspase9) and caspase pathways (Caspase3, 8 and 9) and also by regressing angiogenic factor (VEGF) without any inflammation or adverse side effects. Hence, PAWE serve as a potential antioxidant and antitumor agent.

Keywords: antioxidant, antitumor, Dalton lymphoma ascites (DLA), fungi, Phellinus robinae

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Authors: S. Techaoei, K. Jarmkom, P. Eakwaropas, W. Khobjai

Abstract:

The objective of this research was focused on investigating in vitro antimicrobial activity of Phellinus linteus fruiting body extracts on Pseudomonas aeruginosa, Escherichia coli, Staphylococcus aureus and Methicillin-resistant Staphylococcus aureus. Phellinus linteus fruiting body was extracted with ethanol and ethyl acetate and was vaporized. The disc diffusion assay was used to assess antimicrobial activity against tested bacterial strains. Primary screening of chemical profile of crude extract was determined by using thin layer chromatography. The positive control and the negative control were used as erythromycin and dimethyl sulfoxide, respectively. Initial screening of Phellinus linteus crude extract with the disc diffusion assay demonstrated that only ethanol had greater antimicrobial activity against Pseudomonas aeruginosa, Escherichia coli, Staphylococcus aureus and Methicillin-resistant Staphylococcus aureus. The MIC assay showed that the lower MIC was observed with 0.5 mg/ml of Pseudomonas aeruginosa and Methicillin-resistant Staphylococcus aureus and 0.25 mg/ml. of Escherichia coli and Staphylococcus aureus, respectively. TLC chemical profile of extract was represented at R_f ≈ 0.71-0.76.

Keywords: Staphylococcus aureus, Escherichia coli, Phellinus linteus, Methicillin-resistant Staphylococcus aureus, antimicrobial activity

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Authors: Kiran Nawaz, Ahmad Ali Shahid, Sehrish Iftikhar, Waheed Anwar, Muhammad Nasir Subhani

Abstract:

Chili (Solanum annum L.) attacks by many fungal pathogens, including members of Oomycetes which are responsible for root rot in different chili growing areas of the world. Oomycetes pathogens cause economic losses in different regions of the Pakistan. Most of the plant tissues, including roots, crowns, fruit, and leaves, are vulnerable to Phytophthora capsici. It is very difficult to manage the Phytophthora root rot of chili as many commercial varieties are tremendously vulnerable to P. capsici. The causal agent of the disease was isolated on corn meal agar (CMA) and identified on a morphological basis by using available taxonomic keys. The pathogen was also confirmed on the molecular basis through internal transcribed spacer region and with other molecular markers.The Blastn results showed 100% homology with already reported sequences of P. capsici in NCBI database. Most of the farmers have conventionally relied on foliar fungicide applications to control Phytophthora root rot in spite of their incomplete effectiveness. In this study, in vitro plate assay, seed soaking and foliar applications of 6 fungicides were evaluated against root rot of chili. In vitro assay revealed that significant inhibition of linear growth was obtained with Triflumizole at 7.0%, followed by Thiophanate methyl (8.9%), Etridiazole (6.0%), Propamocarb (5.9%) and 7.5% with Mefenoxam and Iprodione for P. capsici. The promising treatments of in vitro plate bioassay were evaluated in pot experiments under controlled conditions in the greenhouse. All fungicides were applied after at 6-day intervals. Results of pot experiment showed that all treatments considerably inhibited the percentage of P. capsici root rot incidence. In addition, application of seed soaking with all six fungicides combined with the foliar spray of the same components showed the significant reduction in root rot incidence. The combine treatments of all fungicides as in vitro bioassay, seed soaking followed by foliar spray is considered non-harmful control methods which have advantages and limitation. Hence, these applications proved effective and harmless for the management of soil-borne plant pathogens.

Keywords: blastn, bioassay, corn meal agar(CMA), oomycetes

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Authors: Izdihar Ismail, Alona C. Linatoc, Maryati Mohamed

Abstract:

The tropical rainforest of Malaysia is known for its rich biological diversity and high endemicity. The potential for these forests to hold the cure for many diseases and illnesses is high and much is yet to be discovered. This study explores the richness of the tropical rainforest of Endau-Rompin National Park in Johor, Malaysia in search of plants traditionally used by the indigenous people in the treatment of malaria and malaria-like symptoms. Seven species of plants were evaluated and tested for antiplasmodial activities. Different plant parts were subjected to methanolic and aqueous extractions. A total of 24 extracts were evaluated by histidine-rich protein II (HRP2) assay against K1 strain of Plasmodium falciparum chloroquine-resistant. Ten extracts showed significant inhibition of the growth of P. falciparum. Phytochemical screening of the same extracts revealed the presence of alkaloids, flavonoids, terpenoids and anthraquinones. This study affirms that tropical rainforests may still hold undiscovered cures for many diseases and illnesses that have inflicted millions of people worldwide. The species studied herein have not known to have been studied elsewhere before.

Keywords: Endau-Rompin, malaria, Malaysia, tropical rainforest, traditional knowledge

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Authors: McKenzie S. Braley, Lesley Jessiman

Abstract:

If language reveals internal psychological processing, then it is also likely that language use in autobiographical memory transcripts may be used as a window into attachment style and related personality features. The current study, therefore, examined the possible associations between attachment style, negative affectivity, social inhibition, and linguistic features extracted from autobiographical memory transcripts. Young adult participants (n = 61) filled out attachment and personality questionnaires, and orally reported a relationship-related memory. Memories were audio-recorded and later transcribed verbatim. Using a computerized linguistic extraction tool, positive affect words, negative affect words, and cognition words were extracted. Spearman’s rank correlation coefficients revealed that attachment anxiety was negatively correlated with cognition words (r2 = -0.26, p = 0.047) and that negative affectivity was negatively correlated with positive affect words (r2 = -0.32, p = 0.012). The findings suggest that attachment style and personality are associated with speech styles indicative of both emotionality and depth of processing. Because attachment styles, negative affectivity, and social inhibition are associated with poor mental health outcomes, analyses of key linguistics features in autobiographical memory narratives may provide reliable screening tools for mental wellbeing.

Keywords: attachment style, autobiographical memory, language, negative affectivity, social inhibition

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Authors: Pennisi Stefania, Giuffrida Graziella, Coppa Federica, Iannello Giulia, Cartelli Simone, Lo Faro Riccardo, Ferruggia Greta, Brundo Maria Violetta

Abstract:

Tissue engineering aims to develop advanced medical devices to restore normal functions of damaged tissue. These devices, even more effective than conventional methods, are called skin substitutes and are configured as drugs to be applied to the damaged area, to heal extensive and deep wounds which could otherwise lead to chronic wounds lasting over time. Among the variety of commercially available skin substitutes, those that have proven to be most effective are those consisting of a bilayer scaffold. The aim of our research was to design a skin substitute which can promote cell proliferation, cell migration and angiogenesis, and which can guarantee timely closure of the wound with satisfactory aesthetic results, in order to avoid the patient excessive pain, risk of contracting infections and long-term hospitalization. The product was tested in vitro using the Scratch Assay. The assay was carried out both on the matrix modified with hyaluronic acid and on the matrix based only on collagen. In both cases, after 48 hours of exposure the wound scratch was almost completely closed in treated cells compared to untreated control.

Keywords: collagen, hyaluronic acid, scratch- wound-healing assay, tissue regeneration

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Authors: Sulaiman B. Ali Alharbi, Bassam H. Mashat, Naif Abdullah Al-Harbi, Milton Wainwright, Abeer S. Aloufi, Sulamain Alnaimat

Abstract:

Bismuth salicylate was found to inhibit the growth of a range of bacteria and yeast, Candida albican. In general the growth of bacteria did not result in the increase in bismuth solubilisation, in contrast, bismuth solubilisation increased following the growth of C. albicans. A significant increase in the biomass (dry weight) of Aspergillus niger and Aspergillus oryzae occurred in vitro when these fungi were grown in the presence of bismuth salicylate. Biomass increase occurred over a range of bismuth compound additions, which in the case of A. oryzae was associated with the increase in the solubilisation of the insoluble bismuth compounds.

Keywords: bacterial inhibition, fungal growth stimulation, medical uses of bismuth, yeast inhibition

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Authors: HengJiang Cai, RuiJin Zhang, JinSong Gui

Abstract:

Enhanced UV-B (280-320nm) radiation resulting from ozone depletion was one of the global environmental problems. The effects of enhanced UV-B radiation on marine macro-algae were exposed to be the greatest in shallow intertidal environments because the macro-alga was often at or above the water during low tide. Ulva pertusa Kjellman was belonged to Chlorophyta (Phylum), Ulvales (Order), Ulvaceae (Family) which was widely distributed in the western Pacific coast, and the resources were extremely rich in China. Therefore, the effects of UV-B radiation on the growth of Ulva pertusa seedling were studied in this research. Ulva pertusa seedling appearances were mainly characterized by rod shapes and tadpole shapes. The percentage of rod shapes was 90.68%±2.50%. UV-B radiation could inhibit the growth of Ulva pertusa seedling, and the growth inhibition was more significant with the increased doses of UV-B radiation treatment. The relative inhibition rates of Ulva pertusa seedling length were16.11%, 24.98%and 39.04% respectively on the 30th day at different doses (30.96, 61.92 and 123.84 Jm-2d-1) of UV-B radiation. Ulva pertusa seedling had emerged death under UV-B radiation, and the death rates were increased with the increased doses of UV-B radiation treatment. Physiology and biochemistry of Ulva pertusa seedling could be affected by UV-B radiation treatment. The SOD (superoxide dismutase) activity was increased at low-dose UV-B radiation (30.96 Jm-2d-1), while was decreased at high-dose UV-B radiation (61.92 and 123.84 Jm-2d-1). UV-B radiation could inhibit CAT (catalase) activity all the while. It speculated that the reasons for growth inhibition and death of Ulva pertusa seedling were excess ROS (reactive oxygen species), which produced by UV-B radiation.

Keywords: growth, physiology and biochemistry, Ulva pertusa Kjellman, UV-B radiation

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