Search results for: fold changes

Authors: Rubina Ghani, Sehrish Zia, Afifa Fatima Rafique, Shaista Emad

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Background: Cancer is a leading cause of death worldwide, with breast cancer being the most common cancer in women. Pakistan has the highest rate of breast cancer cases among Asian countries. Early and accurate diagnosis is crucial for treatment outcomes and quality of life. Method: It is a case-control study with a sample size of 150. There were 100 suspected cancer cases, 25 healthy controls, and 25 diagnosed cancer cases. To analyze SSAT-1 mRNA expression in whole blood, Zymo Research Quick-RNA Miniprep and Innu SCRIPT—One Step RT-PCR Syber Green kits were used. Patients were divided into three groups: 100 suspected cancer cases, 25 controls, and 25 confirmed breast cancer cases. Result: The total mRNA was isolated, and the expression of SSAT-1 was measured using RT-qPCR. The threshold cycle (Ct) values were used to determine the amount of each mRNA. Ct values were then calculated by taking the difference between the CtSSAT-1 and Ct GAPDH, and further Ct values were calculated with the median absolute deviation for all the samples within the same experimental group. Samples that did not correlate with the results were taken as outliers and excluded from the analysis. The relative fold change is shown as 2^-Ct values. Suspected cases showed a maximum fold change of 32.24, with a control fold change of 1.31. Conclusion: The study reveals an overexpression of SSAT-1 in breast cancer. Furthermore, we can use SSAT-1 as a diagnostic, prognostic, and therapeutic marker for early diagnosis of cancer.

Keywords: breast cancer, spermidine/spermine, qPCR, mRNA

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Authors: Jia-Yu Wu, Chih-Han Chang, Shu-Fen Chuang, Rong-Yang Lai

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Objective: This study was to examine the stress distribution of tooth with different class V restorations under different loading situations and geometry by 3D finite element (FE) analysis. `Methods: A series of FE models of mandibular premolars containing class V cavities were constructed using micro-CT. The class V cavities were assigned as the combinations of different cavity depths x occlusal -gingival heights: 1x2, 1x4, 2x2, and 2x4 mm. Three alveolar bone loss conditions were examined: 0, 1, and 2 mm. 200 N force was exerted on the buccal cusp tip under various directions (vertical, V; obliquely 30° angled, O; oblique and parallel the individual occlusal cavity wall, P). A 3-D FE analysis was performed and the von-Mises stress was used to summarize the data of stress distribution and maximum stress. Results: The maximal stress did not vary in different alveolar bone heights. For each geometry, the maximal stress was found at bilateral corners of the cavity. The peak stress of restorations was significantly higher under load P compared to those under loads V and O while the latter two were similar. 2x2mm cavity exhibited significantly increased (2.88 fold) stress under load P compared to that under load V, followed by 1x2mm (2.11 fold), 2x4mm (1.98 fold) and 1x4mm (1.1fold). Conclusion: Load direction causes the greatest impact on the results of stress, while the effect of alveolar bone loss is minor. Load direction parallel to the cavity wall may enhance the stress concentration especially in deep and narrow class cavities.

Keywords: class v restoration, finite element analysis, loading situation, stress

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Authors: Madhulika Pathak, Polani Seshagiri, Vani Venkatappa

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Blastocyst hatching is an indispensable process for successful implantation. One of the major reasons for implantation failure in IVF clinic is poor quality of embryo, which are not development/hatching-competent. Therefore, attempts are required to develop or enhance the culture system with a molecule recapitulating the autocrine/paracrine factors containing the environment of in-vivo endometrial milieu. We have tried to explore the functional molecules involved in the hamster hatching phenomenon. Blastocyst hatching is governed by several molecules that are entwined and regulate this process, among which cytokines are known to be expressed and are still least explored. Two of such cytokines we have used for our study are IL-1β and its natural antagonist IL-1ra to understand the functional dynamics of cytokines involved in the hatching process. Using hamster, an intriguing experimental model for hatching behavior, we have shown the mRNA (qPCR) and protein (ICC) expression of IL-1β, IL-1ra and IL-1 receptor type 1 throughout all the stages of morula, blastocyst and hatched blastocyst. Post-asserting the expression, the functional role is shown by supplementation studies, where IL-1β supplementation showed enhancement in hatching level (IL-1β treated: 84.1 ± 4.2% vs control: 63.7 ± 3.1 %, N=11), further confirmed by the diminishing effect of IL-1ra on hatching rate (IL-1ra treated: 27.5 ± 11.1% vs control: 67.9 ± 3.1%). The exogenous supplementation of IL-1ra decreased the survival rate of embryos and affected the viability in dose-dependent manner, establishing the importance of IL-1β in blastocyst cell survival. Previously, the cathepsin L and B were established as the proteases that were involved in the hamster hatching process. The inducing effect of IL-1β was correlated with enhanced mRNA level, as analyzed by qPCR, for both CAT L (by 1.9 ± 0.5 fold) and CAT B (by 3.5 ± 0.1) fold which was diminished in presence of IL-1ra (Cat L by 88 percent and Cat B by 94 percent. Moreover, using a specific fluorescent substrate-based assay kit, the enzymatic activity of these proteases was found to be increased in presence of IL-1β (Cat L by 2.1 ± 0.1 fold and CAT B by 2.3 ± 0.7 fold) and was curtailed in presence of IL-1ra. These observations provide functional insights with respect to the involvement of cytokines in the hatching process. This has implications in understanding the hatching biology and improving the embryo development quality in IVF clinics.

Keywords: Blastocyst, Cytokines, Hatching, Interleukin

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Authors: Jasna M. Čanadanović-Brunet, Gordana S. Ćetković, Jelena J. Vulić, Sonja M. Djilas, Vesna T. Tumbas Šaponjac, Sladjana M. Stajčić

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Honey serves as a source of natural antioxidants, which are effective in reducing the risk of heart disease, cancer, immune-system decline, cataracts, different inflammatory processes, and also prevent deteriorative oxidation reactions in foods such as enzymatic browning of fruit and vegetables. Honey is a natural saturated sugar solution, but it also contains certain minor constituents, proteins, enzymes, amino and organic acids, lipids, vitamins, phenolic acids, flavonoids and carotenoids. It is consumed in its natural form alone, but also in combination with nuts and various kinds of dried fruits. The aim of this research was to investigate the contribution of dried cherry on phenols (TPh) and flavonoids (Fl) contents and antioxidant activities of honey. Phenolic compounds in Serbian polyfloral (PH), linden (LH) and acacia (AH) honey and also in their mixtures with dried cherry, in 40% mass concentrations (PH40; LH40, AH40), were determined. In comparison to honey, TPh increased 2.25 times for LH40, 2.16 times for AH40 and 1.45 times for PH40, while Fl increased 2.81-fold for PH40, 1.21-fold for LH40 and 1.44-fold for AH40. Antioxidant activity was investigated with two assays, DPPH test and reducing power (RP), and expressed as EC50DPPH and RP0.5 values. The EC50DPPH values were: EC50PH40 = 1.16 mg/ml; EC50LH40= 1.42 mg/ml and EC50AH40= 1.69 mg/ml, while RP0.5 were: RP0.5PH40 = 15.05 mg/ml; RP0.5LH40 = 16.09 mg/ml and P0.5AH40 = 17.60 mg/ml. Our results indicate that supplementation of polyfloral, linden and acacia honey with 40% dried cherry improves antioxidant activity of honey by enriching the phenolic composition.

Keywords: antioxidant activity, dried cherry, honey, phenolics

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Authors: M. M. M. Jaradat

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For a given graph G, the set Ԑ of all subsets of E(G) forms an |E(G)| dimensional vector space over Z2 with vector addition X⊕Y = (X\Y ) [ (Y \X) and scalar multiplication 1.X = X and 0.X = Ø for all X, Yϵ Ԑ. The cycle space, C(G), of a graph G is the vector subspace of (E; ⊕; .) spanned by the cycles of G. Traditionally there have been two notions of minimality among bases of C(G). First, a basis B of G is called a d-fold if each edge of G occurs in at most d cycles of the basis B. The basis number, b(G), of G is the least non-negative integer d such that C(G) has a d-fold basis; a required basis of C(G) is a basis for which each edge of G belongs to at most b(G) elements of B. Second, a basis B is called a minimum cycle basis (MCB) if its total length Σ BϵB |B| is minimum among all bases of C(G). The lexicographic product GρH has the vertex set V (GρH) = V (G) x V (H) and the edge set E(GρH) = {(u1, v1)(u2, v2)|u1 = u2 and v1 v2 ϵ E(H); or u1u2 ϵ E(G) and there is α ϵ Aut(H) such that α (v1) = v2}. In this work, a construction of a minimum cycle basis for the wreath product of wheels with paths is presented. Also, the length of the longest cycle of a minimum cycle basis is determined. Moreover, the basis number for the wreath product of the same is investigated.

Keywords: cycle space, minimum cycle basis, basis number, wreath product

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Authors: Ossama Abdelmotaal, Olfat Shaker, Tarek Salman, Lamiaa Nabeel, Mostafa Shabayek

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Colorectal Cancer (CRC) is an important worldwide health problem. Colonoscopy is used in detecting CRC suffer from drawbacks where colonoscopy is an invasive method. This study validates easier and less time-consuming techniques to evaluate the usefulness of detecting miRNA-21, miRNA-215 and miRNA-378 in the sera of colorectal cancer patients as new diagnostic tools. This study includes malignant (Colo Rectal Cancer patients, n= 64)) and healthy (n=27) groups. The studied groups were subjected to colonoscopic examination and estimation of miRNA-21, miRNA-215 and miRNA-378 in sera by RT-PCR. miRNA-21 showed the statistically significantly highest median fold change. miRNA-378 showed statistically significantly lower value (Both showed over-expression). miRNA-215 showed the statistically significantly lowest median fold change (It showed down-regulation). Overall the miRNA (21-215 and 378) appear to be promising method of detecting CRC and evaluating its stages.

Keywords: colorectal cancer, miRNA-21, miRNA-215, miRNA-378

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Authors: Marcin Dębowski, Marcin Zieliński, Mirosław Krzemieniewski

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This publication presents a reactor designed for methane fermentation of organic substrates. The design is based on rotating cellular cylinders connected to a biomass feeder and an ultrasonic generator. This allows for simultaneous mixing and partial disintegration of the biomass, as well as stimulating higher metabolic rates within the microorganisms. Such a design allows from 2-fold to 14-fold reduction of power usage when compared to conventional mixing systems. The sludge does not undergo mechanical deformation during the mixing process, which improves substrate biodegradation efficiency by 10-15%. Cavitation occurs near the surface of the rods, partially releasing the biomass and separating it from the destroyed microorganisms. Biogas is released further away from the cellular cylinder rods due to the effect of the ultrasonic waves, in addition to increased biochemical activity of the microorganisms and increased exchange of the nutrient medium with metabolic products, which results in biogas production increase by about 15%.

Keywords: methane fermentation, bioreactors, biomass, mixing system

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Authors: Shirin Jalili, Ramin Ghasemi Shayan

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Coarctation of the aorta (CoA) may be a common (CHD), which is the seventh most common sort of CHD. Still, this is often likely a think little off since the determination may be deferred, indeed within the pediatric populace. The choice for surgical repair incorporates resection of the contracted section with end-to-end or end-to-side anastomosis, subclavian fold aortoplasty, resection, and join the intervention, or prosthetic fix aortoplasty. Drastically expanded end-to-end repair or switched subclavian fold aortoplasty can be utilized when the coarctation expands to the distal arch. Swell angioplasty can be a palliative choice sometime recently the conclusive redress. Its objective is to stabilize high-risk patients that cannot be submitted to quick surgical intercession, such as untimely newborns. For disconnected and discrete coarctations, it can, as a rule, be drawn nearer and repaired by means of cleared out thoracotomy, extraction of the infected aorta (coarctectomy), and remaking, ordinarily by amplified end-to-end anastomosis. In this article, we need to supply a diagram of current proposals and strategies utilized to picture coarctations of the aorta.

Keywords: coarctation of the aorta, congenital heart disease, strategies, surgical repair

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Authors: Khaled Khleifat, Muayyad Abboud, Ahmad Almustafa

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The effect of metabolic inhibitor/uncoupler(s) (CCCP and NaN3) and sulfhydryl reagents (dithiothreitol, 2 mercaptoethanol glutathione) on cadmium uptake was investigated in Enterobacter aerogenes strains. They include a transformed strain bearing the Vitreoscillahemoglobin gene, vgb as well as control strains that lack this transformed gene. The vgb-harboring strains showed better uptake of cadmium than vgb-lacking strains. Under low aeration, there was 2 fold enhancement of Cd+2 uptake in vgb-harboring strains compared with 1.6-fold enhancement under high aeration. The CCCP caused 36, 40 and 58% inhibition in cadmium uptake of parental, pUC9 harboring and VHb expressing cells, respectively. Similarly, the sodium azide exerted 44, 38 and 55% inhibition in Cd+2 uptake of parental, pUC9 harboring and VHb expressing cells, respectively. Less extensive inhibition of Cd+2 uptake in the range of 11 to 39% was observed with sulfhydryl reagents.

Keywords: bacterial hemoglobin, VHb, Cd uptake, biosorption

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Authors: Preshanthan Moodley, E. B. Gueguim-Kana

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The objective of this study was to develop a method to pretreat sugarcane leaf waste using microwave-assisted (MA) inorganic salt. The effects of process parameters of salt concentration, microwave power intensity and pretreatment time on reducing sugar yield from enzymatically hydrolysed sugarcane leaf waste were investigated. Pretreatment models based on MA-NaCl, MA-ZnCl2 and MA-FeCl3 were developed. Maximum reducing sugar yield of 0.406 g/g was obtained with 2 M FeCl3 at 700W for 3.5 min. Scanning electron microscopy (SEM) and Fourier Transform Infrared analysis (FTIR) showed major changes in lignocellulosic structure after MA-FeCl3 pretreatment with 71.5 % hemicellulose solubilization. This pretreatment was further assessed on sorghum leaves and Napier grass under optimal MA-FeCl3 conditions. A 2 fold and 3.1-fold increase in sugar yield respectively were observed compared to previous reports. This pretreatment was highly effective for enhancing enzymatic saccharification of lignocellulosic biomass.

Keywords: acid, pretreatment, salt, sugarcane leaves

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Authors: Kyaw Myint

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Each and everyone seeking happiness. People are trying various kinds of ways to earn happiness. Noble eight fold path is the way to attain the highest peace and happiness which includes three parts Morality ( Sila ) Concentration ( smadi ), and Wisdom ( Pyanna ). According to the Buddha’s teaching to attain higher level of peace and happiness, one must walk on this path . In the Buddha’s teaching morality is basic for all of these three parts. The right Livelihood consider as morality to reach peace or earn happiness in this life and here after next life. It is a way of earn a living without breaking the precepts. The essence of the teaching is to practice metal purity through the material gaining. In this article attempts to study right way of livelihood laid down by the Buddha for lay disciple special reference to Vinijja suttha, Ardiya Sutta, Dighajanu Sutta and singalovoda sutta. This paper approach qualitative research based mainly on documentary analysis. The result of the study shows that right livelihood in Buddha's teaching involves both abstaining from wrong livelihood and taking right livelihood with right view and right effort. Buddha guidance on right livelihood can consist both materially and spiritually for lay disciple.

Keywords: right livelihood, eight fold path, lay disciple, wrong livelihood

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Authors: Rajkumar Ghosh

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Out-of-sequence thrusts (OOST) are reported at different geographic locations with various local names along Siwalik Himalaya (SH), Lesser Himalaya (LH), Higher Himalaya (HH) from Bhutan, India, Nepal, and Pakistan Himalayan range. Most of OOSTs have been identified within the upper LH, and the lower HH based on geochronological age jump across. These thrusts activated from Late Miocene to recent. The Chaura Thrust (CT) was deciphered from age jump of Apatite Fission Track (AFT) and considered as blind thrust base on variable exhumation rates in Chaura region, Satluj river valley, Himachal Pradesh. CT is located north of Jhakri Thrust (JhT) and is also differently identified as Sarahan thrust (ST). Structural documentation from the rocks near the OOST in Chaura was not so far done. Detail structural study of the Jeori Group of rocks was carried out in this study to understand the manifestation of the Chaura thrust and associated structures in meso- to micro-scale. Box fold, scar fold, kink fold, crenulation cleavages, and boudins are developed in the Chaura region. These structures usually do not indicate shear sense. When studied under an optical microscope, the Chaura samples reveal that the mica fish are usually lenticular with aspect ratio (R) varying from 6–11 and inclination angle (α) from 15–40°. According to ‘R’ and ‘α’, elongated sigmoid shaped mica fish and parallelogram shaped mica fish were also documented. Asymmetric mica fish demonstrate top-to-S/SW ductile shear, which is similar as that of Chaura thrust. Grain boundary migration (GBM) structures in quartzo-feldspathic grains from Jeori Group of rocks indicate deformation temperature ranging from 400 to 650°C. This can indicate that the OOST at Chaura, i.e., the Chaura Thrust, underwent thrusting in the ductile regime.

Keywords: out-of-sequence thrust, chaura thrust, sarahan thrust, jakhri thrust, higher himalaya, s/c- fabric

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Authors: Tatiana R. Simonyan, Elena A. Protasova, Anastasia V. Mamontova, Eugene G. Maksimov, Konstantin A. Lukyanov, Alexey M. Bogdanov

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Here, we describe two variants of the calcium indicators based on the GCaMP sensitive core and BrUSLEE fluorescent protein (GCaMP-BrUSLEE and GCaMP-BrUSLEE-145). In contrast to the conventional GCaMP6-family indicators, these fluorophores are characterized by the well-marked responsiveness of their fluorescence decay kinetics to external calcium concentration both in vitro and in cellulo. Specifically, we show that the purified GCaMP-BrUSLEE and GCaMP-BrUSLEE-145 exhibit three-component fluorescence decay kinetics, with the amplitude-normalized lifetime component (t3*A3) of GCaMP-BrUSLEE-145 changing four-fold (500-2000 a.u.) in response to a Ca²⁺ concentration shift in the range of 0—350 nM. Time-resolved fluorescence microscopy of live cells displays the two-fold change of the GCaMP-BrUSLEE-145 mean lifetime upon histamine-stimulated calcium release. The aforementioned Ca²⁺-dependence calls considering the GCaMP-BrUSLEE-145 as a prospective Ca²⁺-indicator with the signal read-out in the time domain.

Keywords: calcium imaging, fluorescence lifetime imaging microscopy, fluorescent proteins, genetically encoded indicators

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Authors: Massimo Mozzon, Nadia Raffaelli

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Proteases from herbs, woody plants, and trees are exploited for cheesemaking in several countries, especially in South Europe and West Africa. Particularly, “thistles” belonging to several genera within the Asteraceae family (Cynara, Silybum, Centaurea, Carlina, Cirsium, Onopordum) are traditionally used in Mediterranean countries for clotting raw ewe’s and goat’s milk. For the first time, the clotting performance of an aqueous extract from flowers of Onopordum tauricum Willd. (Taurian thistle, bull cottonthistle) were tested in milk of different origin (cow, goat, ewe). The vegetable material was collected in the Central Apennines range, between the Marche and Umbria regions. A response surface methodology (RSM) approach was used to study the effect of the curdling variables (temperature, pH, amount of enzymatic extract) on the technological performance of the thistle extract. A three-step procedure for the purification of the enzyme (ammonium sulphate precipitation, gel filtration and ion-exchange chromatography) was also carried out. The milk clotting activity (MCA) of O. tauricum crude extracts was strongly affected by temperature, pH and by the interaction between these two variables, according to a second-order response surface model, while the milk/coagulant ratio did not affect in a significant way the clotting properties. Experimental data showed that the addition of 10 mM CaCl2 reduced the clotting time of ewe’s, goat’s, and cow’s milk by about 3-fold, 8-fold, and 14-fold, respectively, at 35°C and pH 6.7-6.8. After purification, an enzymatic preparation very close to homogeneity was obtained, which showed a major band at about 30 kDa when analyzed by SDS-PAGE. The identity of the enzyme as an aspartic protease was confirmed by inhibition studies. Cheese-making trials were carried out to check the scale-up (1 to 5 L of milk; 37 °C; 10 mM CaCl2 fortification) and set the recipe: 35-45% of curd yields were recorded, according to curd cutting and pressing.

Keywords: milk clotting activity, Onopordum tauricum, plant proteases, vegetable rennet

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Authors: Dalia G. Aseel

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The potato (Solanum tubersum, L.) has become one of the major vegetable crops in Egypt and all over the world. Potato leafroll virus(PLRV) was observed on potato plants collected from different governorates in Egypt. Three cultivars, Spunta, Diamont, and Cara, infected with PLRV were collected; RNA was extracted and subjected to Real-Time PCR using the coat protein gene primers. The results showed that the expression of the coat protein was 39.6-fold, 12.45-fold, and 47.43-fold, respectively, for Spunta, Diamont, and Cara cultivars. Differential Display Polymerase Chain Reaction (DD-PCR) using pathogenesis-related protein 1 (PR-1), β-1,3-glucanases (PR-2), chitinase (PR-3), peroxidase (POD), and polyphenol oxidase (PPO) forward primers for pathogenesis-related proteins (PR). The obtained data revealed different banding patterns depending on the viral type and the region of infection. Regarding PLRV, 58 up-regulated and 19 down-regulated genes were detected. Sequence analysis of the up-and down-regulated genes revealed that infected plants were observed in comparison with the healthy control. Sequence analysis of the up-regulated gene was performed, and the encoding sequence analysis showed that the obtained genes include: induced stolen tip protein. On the other hand, two down-regulated genes were identified: disease resistance RPP-like protein and non-specific lipid-transfer protein. In this study, the expressions of PR-1, PR-2, PR-3, POD, and PPO genes in the infected leaves of three potato cultivars were estimated by quantitative real-time PCR. We can conclude that the PLRV-infection of potato plants inhibited the expression of the five PR genes. On the contrary, infected leaves by PLRV elevated the expression of some defense genes. This interaction may also induce and/or suppress the expression of some genes responsible for the plant's defense mechanisms.

Keywords: PLRV, pathogenesis-related proteins (PRs), DD-PCR, sequence, real-time PCR

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Authors: Vongsathorn Ngampuak, Yutachai Chookaew, Wipawee Dejtisakdi

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Dunaliella salina has great potential as a system for generating commercially valuable products, including beta-carotene, pharmaceuticals, and biofuels. Our goal is to improve this potential by enhancing growth rate and other properties of D. salina under optimal growth conditions. We used ethyl methane sulfonate (EMS) to generate random mutants in D. salina KU11, a strain classified in Thailand. In a preliminary experiment, we first treated D. salina cells with 0%, 0.8%, 1.0%, 1.2%, 1.44% and 1.66% EMS to generate a killing curve. After that, we randomly picked 30 candidates from approximately 300 isolated survivor colonies from the 1.44% EMS treatment (which permitted 30% survival) as an initial test of the mutant screen. Among the 30 survivor lines, we found that 2 strains (mutant #17 and #24) had significantly improved growth rates and cell number accumulation at stationary phase approximately up to 1.8 and 1.45 fold, respectively, 2 strains (mutant #6 and #23) had significantly decreased growth rates and cell number accumulation at stationary phase approximately down to 1.4 and 1.35 fold, respectively, while 26 of 30 lines had similar growth rates compared with the wild type control. We also analyzed cell size for each strain and found there was no significant difference comparing all mutants with the wild type. In addition, mutant #24 had shown an increase of biomass accumulation approximately 1.65 fold compared with the wild type strain on day 5 that was entering early stationary phase. From these preliminary results, it could be feasible to identify D. salina mutants with significant improved growth rate, cell accumulation and biomass production compared to the wild type for the further study; this makes it possible to improve this microorganism as a platform for biotechnology application.

Keywords: Dunaliella salina, ethyl methyl sulfonate, growth rate, biomass

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Authors: Jae Woong Sull

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Lipid levels are related to the risk of cardiovascular diseases. Cholesterol ester transfer protein (CETP) gene is one of the candidate genes of cardiovascular diseases. A total of 2,304 persons were chosen from a Hospital (N=4,294) in South Korea. Female subjects with the CG/GG genotype had a 2.03 -fold (p=0.0001) higher risk of having abnormal HDL cholesterol levels (<40 mg/dL) than subjects with the CC genotype. Male subjects with the CG/GG genotype had a 1.34 -fold (p=0.0019) higher risk than subjects with the CC genotype. When analyzed by body mass index, the association with CETP was much stronger in male subjects with BMI>=25.69 (OR=1.55, 95% CI: 1.15-2.07, P=0.0037) than in male lean subjects. When analyzed by family history of diabetes, the association with CETP was much stronger in male subjects with positive family history of low physical activity (OR=4.82, 95% CI: 1.86-12.5, P=0.0012) than in male subjects with negative family history of diabetes. This study clearly demonstrates that genetic variants in CETP influence HDL cholesterol levels in Korean adults.

Keywords: CETP, diabetes, obesity, polymorphisms

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Authors: Ennus Tajuddin Tamboli, Madhukar Garg, Mohd. Mujeeb, Sayeed Ahmad

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An efficient protocol for simultaneous production of forskolin and rosmarinic acid in in vitro callus derived from the leaves of Coleus forskohlii Briq. has been developed. MS media was used for the establishment of cultures and NAA + 6-BA (1.0 ppm) was found best for callus growth. The callus was further subjected to treatment with various elicitor/precursors viz. chitosan, thidiazuron and methyl jasmonate to observe their effect on production of biomass and accumulation of secondary metabolites. The content of forskolin and rosmarinic acid were estimated by HPTLC, in comparison to natural explant which showed 2 fold and 10 fold rise in forskolin and rosmarinic acid content, respectively. Methy1 jasmonate 50 µM was found best for production of forskolin, whereas thidiazuron showed best results in the yield of rosmarinic acid, separately in static culture. However, combined treatment in suspension culture showed moderated effect for increase in secondary metabolites but the biomass increased significantly as compared to static culture.

Keywords: plant tissue culture, secondary metabolites, coleus, forskolin, rosmarinic acid, HPTLC

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Authors: Weili Shao, Qian Wang, Jianxin He

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Recently, the applications of graphene oxide in fabricating scaffolds for bone tissue engineering have been received extensive concern. In this work, poly l-lactic/graphene oxide composite nanofibers were successfully fabricated by electrospinning. The morphology structure, porosity and mechanical properties of the composite nanofibers were characterized using different techniques. And mouse mesenchymal stem cells were cultured on the composite nanofiber scaffolds to assess their suitability for bone tissue engineering. The results indicated that the composite nanofiber scaffolds had finer fiber diameter and higher porosity as compared with pure poly l-lactic nanofibers. Furthermore, incorporation of graphene oxide into the poly l-lactic nanofibers increased protein adsorptivity, boosted the Young’s modulus and tensile strength by nearly 4.2-fold and 3.5-fold, respectively, and significantly enhanced adhesion, proliferation, and osteogenesis in mouse mesenchymal stem cells. The results indicate that composite nanofibers could be excellent and versatile scaffolds for bone tissue engineering.

Keywords: poly l-lactic, graphene oxide, osteogenesis, bone tissue engineering

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Authors: Jae Woong Sull, Sun Ha Jee

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High-density lipoprotein (HDL) cholesterol levels are associated with decreased risk of coronary artery disease. Several genome-wide association studies (GWAS) for HDL cholesterol levels have implicated cholesterol ester transfer protein (CETP) as possibly causal. We tested for the association between single nucleotide polymorphisms (SNPs) in CETP gene and HDL cholesterol levels in Korean population. Subjects were selected from the Korean Metabolic Syndrome Research Initiative study in the Bundang-Gu area. A total of 2,304 individuals from Bundang-Gu were recruited in 2008. Other subjects were selected from the Severance Hospital (N=4,294). SNP rs6499861 in the CETP gene was associated with mean HDL cholesterol levels (effect per allele -2.044 mg/dL, p=7.23×10-7). Subjects with the CG/GG genotype had a 1.46 -fold (range 1.24–1.72-fold) higher risk of having abnormal HDL cholesterol levels (<40 mg/dL) than subjects with the CC genotype. When analyzed by gender, the association of CETP was stronger in women than in men. When analyzed by physical activity behavior, the association with CETP was much stronger in male subjects with low physical activity (OR=1.54, 95% CI: 1.23-1.92, P=0.0001) than in male subjects with high physical activity. This study clearly demonstrates that genetic variants in CETP influence HDL cholesterol levels in Korean adults.

Keywords: CETP, HDL cholesterol, physical activity, polymorphisms

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Authors: Prativa Das, Lay Poh Tan

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Three-dimensional microenvironment is a need to study the event cascades of liver fibrosis in vitro. Electrospun nanofibers modified with essential extracellular matrix proteins can closely mimic the random fibrous structure of native liver extracellular matrix (ECM). In this study, we fabricate a series of 3D electrospun scaffolds by wet electrospinning process modified with different ratios of collagen-I to fibronectin to achieve optimized distribution of these two ECM proteins on the fiber surface. A ratio of 3:1 of collagen-I to fibronectin was found to be optimum for surface modification of electrospun poly(lactic-co-glycolic acid) (PLGA) fibers by chemisorption process. In 3:1 collagen-I to fibronectin modified scaffolds the total protein content increased by ~2 fold compared to collagen-I modified and ~1.5 fold compared to 1:1/9:1 collagen-I to fibronectin modified scaffolds. We have cultured LX-2 cells on this scaffold over 14 days and found that LX-2 cells acquired more quiescent phenotype throughout the culture period and shown significantly lower expression of alpha smooth muscle actin and collagen-I. Thus, this system can be used as a model to study liver fibrosis by using different fibrogenic mediators in vitro.

Keywords: electrospinning, collagen-I and fibronectin, surface modification of fiber, LX-2 cells, liver fibrosis

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Authors: B. Gepts, E. Meex, E. Nuyts, E. Knaepen, G. Verbeeck

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The importance of resource efficiency and environmental impact assessment has raised the interest in knowing the amount of materials used in buildings. If no BIM model or energy performance certificate is available, material quantities can be obtained through an estimation or time-consuming calculation. For the interior wall area, no validated estimation method exists. However, in the case of environmental impact assessment or evaluating the existing building stock as future material banks, knowledge of the material quantities used in interior walls is indispensable. This paper presents a validated method for the estimation of the interior wall area for dwellings based on easy-to-collect building characteristics. A database of 4963 residential buildings spread all over Belgium is used. The data are collected through onsite measurements of the buildings during the construction phase (between mid-2010 and mid-2017). The interior wall area refers to the area of all interior walls in the building, including the inner leaf of exterior (party) walls, minus the area of windows and doors, unless mentioned otherwise. The two predictive modelling techniques used are 1) a (stepwise) linear regression and 2) a decision tree. The best estimation method is selected based on the best R² k-fold (5) fit. The research shows that the building volume is by far the most important variable to estimate the interior wall area. A stepwise regression based on building volume per building, building typology, and type of house provides the best fit, with R² k-fold (5) = 0.88. Although the best R² k-fold value is obtained when the other parameters ‘building typology’ and ‘type of house’ are included, the contribution of these variables can be seen as statistically significant but practically irrelevant. Thus, if these parameters are not available, a simplified estimation method based on only the volume of the building can also be applied (R² k-fold = 0.87). The robustness and precision of the method (output) are validated three times. Firstly, the prediction of the interior wall area is checked by means of alternative calculations of the building volume and of the interior wall area; thus, other definitions are applied to the same data. Secondly, the output is tested on an extension of the database, so it has the same definitions but on other data. Thirdly, the output is checked on an unrelated database with other definitions and other data. The validation of the estimation methods demonstrates that the methods remain accurate when underlying data are changed. The method can support environmental as well as economic dimensions of impact assessment, as it can be used in early design. As it allows the prediction of the amount of interior wall materials to be produced in the future or that might become available after demolition, the presented estimation method can be part of material flow analyses on input and on output.

Keywords: buildings as material banks, building stock, estimation method, interior wall area

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Authors: Aymen Arfaoui, Abdelkader Soumaya, Noureddine Ben Ayed

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The Rouas-Ruissateis a part of TunisianAtlas system. Analyze of the collected field data allowed us to propose a new interpretation for the main structural features of thisregion. Tectonic inversions along NE-SW trending fault of Zaghouan and holokinetic movements are the main factors controlling the architecture and geometry of the Jebel Rouas-Ruissate. The presence of breccias, Slumps, and synsedimentaryfaults along NW-SE and N-S trending major faults show that they were active during the Mesozoicextensionalepisodes. During Cenozoic inversion period, this structurewas shaped as imbricatefansformed byNE-SW trending thrust faults. The angularunconformitybetweenupperEocene- Oligocene, and Cretaceousdeposits reveals a compressive Eocene tectonic phase (called Pyrenean phase)occurred duringPaleocene-lower Eocene.The Triassicsaltsacted as a decollementlevel in the NE-SW trendingfault propagation fold model of the Rouas-Ruissate.The inversion of fault-slip data along the main regional fault zones reveals a coexistence of strike-slip and reverse fault stress regimes with NW-SE maximum horizontal stress(SHmax) characterizing the Alpine compressive phase (Upper Tortonian).

Keywords: tunisia, imbricate fans, triassic decollement level, fault propagation fold

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Authors: Nino Kobakhidze, Endi Varsimashvili, Davit Makadze

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The Caucasus is a link of the Alpine-Himalayan fold belt and involves the Greater Caucasus and the Lesser Caucasus fold systems and the Intermountain area. The study object is located within the northernmost part of the Lesser Caucasus orogen, in the eastern part of Achara-Trialeti fold -thrust belt. This area was rather well surveyed in 70th of the twentieth century in terms of oil-and-gas potential, but to our best knowledge, detailed sedimentological studies have not been conducted so far. In order to fill this gap, the authors of the present thesis started research in this direction. One of the objects selected for the research was the deposits of the Kavtura river valley situated on the northern slope of the Trialeti ridge. Paleocene-Lower Eocene deposits known in scientific literature as ‘Borjomi Flysch’ (Turbidites) are exposed in the mentioned area. During the research, the following methodologies were applied: selection of key cross sections, a collection of rock samples, microscopic description of thin sections, mineralogical and petrological analysis of material and identification of trace fossils. The study of Paleocene-Lower Eocene deposits starts with Kavtura river valley in the east, where they are well characterized by microfauna. The cross-section of the deposits starts with Danian variegated marlstone conformably overlain by the alternation of thick and thin-bedded sandstones (thickness 40-50 cm). They are continued with interbedded of thin-bedded sandstones and shales(thickness 4-5 m). On the sole surface of sandstones ichnogenera ‘Helmintopsis’ and ‘Scolicia’ are recorded and within the bed –‘Chondrites’ is found. Towards the Riverhead, there is a 1-2 m gap in sedimentation; then again the Paleocene-Lower Eocene sediments crop out. They starting with alternation of grey-green medium-grained sandstones and shales enclosing dark color plant detritus. They are overlain by the interbedded of calcareous sandstones and marls, where the thickness of sandstones is variable (20-70 cm). Ichnogenus – ‘Scolicia’ is found here. Upwards the above-mentioned deposits pass into Middle Eocenian volcanogenic-sedimentary suits. In the Kavtura river valley, the thickness of the Paleocene-Lower Eocene deposits is 300-400 m. In the process of research, the following activities are conducted: the facial analysis of host rocks, correlation of the study section with other cross sections and interpretation of depositional environment of the area. In the area the authors have found and described ichnogenera; their preliminary determination have shown that they belong to pre-depositional (‘Helmintopsis’) and post-depositional (‘Chondrites’) forms. As known, during the Cretaceous-Paleogene time, the Achara-Trialeti fold-thrust belt extensional basin was the accumulation area with great thicknesses (from shallow to deep marine sediments). It is confirmed once more by the authors investigations preliminary results of paleoichnological studies inclusive.

Keywords: flysh deposits, lithology, The Lesser Caucasus, trace fossils

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Authors: Dragos Dragomir-Stanciu, Leon Marsh

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Background As a response to the gaps identified in recent research in the provision of residential care to address co-occurring health needs, including mental health problems and complexities Gamble Aware has facilitated the possibility to provide a new service which would extend the NGTS provision of residential rehabilitation for gambling disorder with complex and co-morbid presentation. Gordon Moody, together with Adferiad have been successful in securing the tender for this service and this presentation aims to introduce FOLD, the resulting model of treatment developed for the delivery of the service. Setting As a partnership, we have come together to coproduce a model which allows us to share our clinical and industry knowledge and build on our reputations as trusted treatment providers. The presentation will outline our expertise share in development of a unified approach to recovery-oriented models of care, clinical governance, risk assessment and management and aftercare and continuous recovery. We will also introduce our innovative specialist referral portal which will offer referring partners the ability to include the service user in planning their own recovery journey. Outcomes Our collaboration has resulted in the development of the FOLD model which includes three agile and flexible treatment packages aimed at offering the most enhanced and comprehensive treatment in UK, to date, for those most affected by gambling harm. The paper will offer insight into each treatment package and all recovery model stages involved, as well as into the partnership work with NGST providers, local mental health and social care providers and lived experience organisation that will enable us to offer support to more 100 people a year who would otherwise get “lost in the system”. Conclusion FOLD offers a great opportunity to develop, implement and evaluate a new, much needed, whole-person and whole-system approach to counter gambling related harms.

Keywords: gambling treatment, partnership working, integrated care pathways, NGTS, complex needs

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Authors: Pelin Balcik Ercin

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Introduction: ZEB transcription factor family member ZEB2, has a role in epithelial to mesenchymal transition during development and metastasis. The altered circulating extracellular miRNAs expression is observed in diseases, and extracellular miRNAs have an important role in cancer cell microenvironment. In ChIP-Seq study, the expression of miR-2110 was found to be regulated by ZEB2. In this study, the effects of miR2110 on cell proliferation and migration of hepatocellular carcinoma (HCC) cells were examined. Material and Methods: SNU398 cells transfected with mimic miR2110 (20nM) (HMI0375, Sigma-Aldrich) and negative control miR (HMC0002, Sigma-Aldrich). MicroRNA isolation was accomplished with miRVANA isolation kit according to manufacturer instructions. cDNA synthesis was performed expression, respectively, and calibrated with Ct of controls. The real-time quantitative PCR (RT-qPCR) reaction was performed using the TaqMan Fast Advanced Master Mix (Thermo Sci.). Ct values of miR2110 were normalized to miR-186-5p and miR16-5p for the intracellular gene. Cell proliferation analysis was analyzed with the xCELLigence RTCA System. Wound healing assay was analyzed with the ImageJ program and relative fold change calculated. Results: The mimic-miR-2110 transfected SNU398 cells nearly nine-fold (log2) more miR-2110 expressed compared to negative control transfected cells. The mimic-miR-2110 transfected HCC cell proliferation significantly inhibited compared to the negative control cells. Furthermore, miR-2110-SNU398 cell migration capacity was relatively four-fold decreased compared to negative control-miR-SNU398 cells. Conclusion: Our results suggest the miR-2110 inhibited cell proliferation and also miR-2110 negatively affect cell migration compared to control groups in HCC cells. These data suggest the complexity of microRNA EMT transcription factors regulation. These initial results are pointed out the predictive biomarker capacity of miR-2110 in HCC.

Keywords: epithelial to mesenchymal transition, EMT, hepatocellular carcinoma cells, micro-RNA-2110, ZEB2

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Authors: Palaniyandi Karuppaiya, Hsin Sheng Tsay, Fang Chen

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Medicinal herbs do represent a huge and noteworthy reservoir for novel anticancer drugs discovery. Dysosma pleiantha (Hance) Woodson (Berberidaceae), one of the oldest traditional Chinese medicinal herb, highly prized by the mountain tribes of Taiwan and China for its medicinal properties contained pharmaceutically important antitumor compounds podophyllotoxin, quercetin and kaempferol. Among lignans, podophyllotoxin is an active antitumor compound and has now been modified to produce clinically useful drugs etoposide and teniposide. In recent years, natural populations of D. peliantha have declined considerably due to anthropogenic activities such as habitat destruction and commercial exploitation for medicinal applications. As to its overall conservation status, D. pleiantha has been ranked as threatened on the China Species Red List. In the present study, an efficient in vitro callus culture system of D. pleiantha was established on Gamborg’s medium with various combinations and concentrations of different auxins and cytokinins under dark condition. Best callus induction was recorded in 2 mg/L 2, 4 - Dichlorophenoxyacetic acid (2,4-D) along with 0.2 mg/L kinetin and the maximum callus proliferation was achieved at 1 mg/L 2,4-D. Among the explants tested, maximum callus induction (86 %) was achieved from tender leaves. Hence, in subsequent experiments, leaf callus was further investigated for suitable callus biomass and production level of anticancer compounds under the influence of different additives. A maximum fresh callus biomass (8.765 g) was recorded in callus proliferation medium contained 500 mg/L casein hydrolysate. High performance liquid chromatography results revealed that the addition of different concentrations of peptone (1, 2 and 4 g/L) in callus proliferation medium enhanced podophyllotoxin (16 fold), quercetin (12 fold) and kaempferol (5 fold) accumulation than control. Thus, the established in vitro callus culture under the influence of different additives may offer an alternative source of enhanced production of podophyllotoxin, kaempferol and quecertin without harming natural plant population.

Keywords: dysosma pleiantha, kaempferol, podophyllotoxin, quercetin

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Authors: Harika Atmaca, Emir Bozkurt, Latife Merve Oktay, Selim Uzunoglu, Ruchan Uslu, Burçak Karaca

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Microarrays have been developed for highly parallel enzyme-linked immunosorbent assay (ELISA) applications. The most common protein arrays are produced by using multiple monoclonal antibodies, since they are robust molecules which can be easily handled and immobilized by standard procedures without loss of activity. Protein expression profiling with protein array technology allows simultaneous analysis of the protein expression pattern of a large number of proteins. Trabectedin, a tetrahydroisoquinoline alkaloid derived from a Caribbean tunicate, Ecteinascidia turbinata, has been shown to have antitumor effects. Here, we used a novel proteomic approach to explore the mechanism of action of trabectedin in prostate cancer cell line PC-3 by apoptosis antibody microarray. XTT cell proliferation kit and Cell Death Detection Elisa Plus Kit (Roche) was used for measuring cytotoxicity and apoptosis. Human Apoptosis Protein Array (R&D Systems) which consists of 35 apoptosis related proteins was used to assess the omic protein expression pattern. Trabectedin induced cytotoxicity and apoptosis in prostate cancer cells in a time and concentration-dependent manner. The expression levels of the death receptor pathway molecules, TRAIL-R1/DR4, TRAIL R2/DR5, TNF R1/TNFRSF1A, FADD were significantly increased by 4.0-, 21.0-, 4.20- and 11.5-fold by trabectedin treatment in PC-3 cells. Moreover, mitochondrial pathway related pro-apoptotic proteins Bax, Bad, Cytochrome c, and Cleaved Caspase-3 expressions were induced by 2.68-, 2.07-, 2.8-, and 4.5-fold and the expression levels of anti-apoptotic proteins Bcl-2 and Bcl-XL were reduced by 3.5- and 5.2-fold in PC-3 cells. Proteomic (antibody microarray) analysis suggests that the mechanism of action of trabectedin may be exerted via the induction of both intrinsic and extrinsic apoptotic pathways. The antibody microarray platform can be utilised to explore the molecular mechanism of action of novel anticancer agents.

Keywords: trabectedin, prostate cancer, omic protein expression profile, apoptosis

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Authors: Cuneyt Yucelbas, Seral Ozsen, Sule Yucelbas, Gulay Tezel

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Due to the fact that there exist only a small number of complex systems in artificial immune system (AIS) that work out nonlinear problems, nonlinear AIS approaches, among the well-known solution techniques, need to be developed. Gaussian function is usually used as similarity estimation in classification problems and pattern recognition. In this study, diagnosis of breast cancer, the second type of the most widespread cancer in women, was performed with different distance calculation functions that euclidean, gaussian and gaussian-euclidean hybrid function in the clonal selection model of classical AIS on Wisconsin Breast Cancer Dataset (WBCD), which was taken from the University of California, Irvine Machine-Learning Repository. We used 3-fold cross validation method to train and test the dataset. According to the results, the maximum test classification accuracy was reported as 97.35% by using of gaussian-euclidean hybrid function for fold-3. Also, mean of test classification accuracies for all of functions were obtained as 94.78%, 94.45% and 95.31% with use of euclidean, gaussian and gaussian-euclidean, respectively. With these results, gaussian-euclidean hybrid function seems to be a potential distance calculation method, and it may be considered as an alternative distance calculation method for hard nonlinear classification problems.

Keywords: artificial immune system, breast cancer diagnosis, Euclidean function, Gaussian function

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Authors: Vinod Kumar

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Background: Tissue Plasminogen Activator (tPA) showed a level 1 benefit in acute stroke (within 3-6 hrs). Intracarotid sodium nitroprusside (ICSNP) has been studied in this context with a wide treatment window, fast recovery and affordability. This work proposes two mechanisms for acute cases and one mechanism for chronic cases, which are interrelated, for physiological recovery. a)Retrograde Neurotransmission (acute cases): 1)Normal excitatory impulse: at the synaptic level, glutamate activates NMDA receptors, with nitric oxide synthetase (NOS) on the postsynaptic membrane, for further propagation by the calcium-calmodulin complex. Nitric oxide (NO, produced by NOS) travels backward across the chemical synapse and binds the axon-terminal NO receptor/sGC of a presynaptic neuron, regulating anterograde neurotransmission (ANT) via retrograde neurotransmission (RNT). Heme is the ligand-binding site of the NO receptor/sGC. Heme exhibits > 10,000-fold higher affinity for NO than for oxygen (the 10,000-fold effect) and is completed in 20 msec. 2)Pathological conditions: normal synaptic activity, including both ANT and RNT, is absent. A NO donor (SNP) releases NO from NOS in the postsynaptic region. NO travels backward across a chemical synapse to bind to the heme of a NO receptor in the axon terminal of a presynaptic neuron, generating an impulse, as under normal conditions. b)Vasospasm: (acute cases) Perforators show vasospastic activity. NO vasodilates the perforators via the NO-cAMP pathway. c)Long-Term Potentıatıon (LTP): (chronic cases) The NO–cGMP-pathway plays a role in LTP at many synapses throughout the CNS and at the neuromuscular junction. LTP has been reviewed both generally and with respect to brain regions specific for memory/learning. Aims/Study Des’gn: The principles of “generation of impulses from the presynaptic region to the postsynaptic region by very potent RNT (10,000-fold effect)” and “vasodilation of arteriolar perforators” are the basis of the authors’ hypothesis to treat stroke cases. Case-control prospective study. Mater’als And Methods: The experimental population included 82 stroke patients (10 patients were given control treatments without superfusion or with 5% dextrose superfusion, and 72 patients comprised the ICSNP group). The mean time for superfusion was 9.5 days post-stroke. Pre- and post-ICSNP status was monitored by NIHSS, MRI and TCD. Results: After 90 seconds in the ICSNP group, the mean change in the NIHSS score was a decrease of 1.44 points, or 6.55%; after 2 h, there was a decrease of 1.16 points; after 24 h, there was an increase of 0.66 points, 2.25%, compared to the control-group increase of 0.7 points, or 3.53%; at 7 days, there was an 8.61-point decrease, 44.58%, compared to the control-group increase of 2.55 points, or 22.37%; at 2 months in ICSNP, there was a 6.94-points decrease, 62.80%, compared to the control-group decrease of 2.77 points, or 8.78%. TCD was documented and improvements were noted. Conclusions: ICSNP is a swift-acting drug in the treatment of stroke, acting within 90 seconds on day 9.5 post-stroke with a small decrease after 24 hours. The drug recovers from this decrease quickly.

Keywords: brain infarcts, intracarotid sodium nitroprusside, perforators, vasodilatıons, retrograde transmission, the 10, 000-fold effect

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