Macrobrachium nipponense. The full-length cDNA of MnHc-1 was 2,163 bp with a 2,028-bp open reading frame (ORF) encoding a polypeptide of 675 amino acids. The MnHc-1 mRNA was expressed in the hepatopancreas, gill, hemocytes, intestine, ovary, and stomach, with the highest level in the hepatopancreas. In the infection trial, the MnHc-1 mRNA transcripts in the hemocytes were significantly downregulated at 3 h after injection of Aeromonas hydrophila and then upregulated at 6 h and 12 h, followed by a gradual recovery from 24 to 48 h. The MnHc-1 transcriptional expression in the hepatopancreas was measured after M. nipponense were fed seven diets with 2.8, 12.2, 20.9, 29.8, 43.1, 78.9, and 157.1 mg Cu kg−1 for 8 weeks, respectively. The level of MnHc-1 mRNA was significantly higher in the prawns fed 43.1–157.1 mg Cu kg−1 diet than in that fed 2.8–29.8 mg Cu kg−1 diet. This study indicated that the MnHc-1 expression can be affected by dietary copper and the hemocyanin may potentially participate in the antibacterial defense of M. nipponense.Hemocyanin is a copper-containing protein with immune function against disease. In this study, a hemocyanin subunit named MnHc-1 was cloned from Litopenaeus vannamei [ Eriocheir sinensis [ Cherax quadricarinatus [ Homarus americanus [ Caridina multidentata and Atyopsis moluccensis [Crustaceans like other invertebrates only have innate immunity, including many immune molecules to eliminate exogenous pathogens . To datevannamei , Eriochesinensis , Cherax arinatus , Homarusericanus , and Caruccensis . Penaeus monodon [ E. sinensis [ E. sinensis was affected by dietary copper level [Copper is a central component of hemocyanin and a cofactor of many other enzymes, like superoxide dismutase, cytochrome oxidase, and lysyl oxidase , 20. The monodon  and E. ssinensis . It was er level . InvestiMacrobrachium nipponense) is an important aquaculture species in China and other Southeast Asian countries [ M. nipponense farming population due to intensive culture and environmental pollution [ Aeromonas hydrophila is one of the major diseases which can cause 30% death of prawn, sometimes as high as 70% [ M. nipponense innate immune mechanism against A. hydrophila has become a key issue of health management in crustacean farming. The hemocyanin subunit is a functional group for crustacean immunity, and a hemocyanin subunit has been cloned and characterized in the freshwater prawn M. nipponense [ M. nipponense (MnHc-1). To understand the role of MnHc-1 in M. nipponense immunization, (1) the full-length cDNAs of MnHc-1 was cloned, (2) the distribution of MnHc-1 in different tissues was examined, (3) the mRNA expression of MnHc-1 in immune defense after prawn challenge with A. hydrophila was examined, and (4) the relative expression level of MnHc-1 in the hepatopancreas of M. nipponense fed different levels of copper was quantified.Oriental river prawn (ountries . Variousollution . For exah as 70% . As a repponense . In thisM. nipponense) were obtained from a local farm in Shanghai. The hepatopancreas, muscle, gill, ovary, intestine, heart, and stomach were collected from healthy prawn, flash-frozen in liquid nitrogen, and stored at −80°C until RNA extraction. A total of 1 mL hemolymph was collected from the ventral sinus using a sterile syringe and diluted using half volume of anticoagulant solution [All adult oriental river prawns  obtained from Shanghai Ocean University, while each prawn in the control group received the same volume of saline injection. After injection, prawns were put back to the rearing tanks, and hemocyte samples were collected at 0, 3, 6, 12, 24, and 48 h after injection, centrifuged, and stored at −80°C for RNA extraction.Prior to the challenge experiment, the adult prawns were acclimatized in the laboratory for 2 weeks. A total of 300 healthy prawns were randomly divided into two groups with five replicates. According to the preliminary experiment, the prawn in the bacterial challenge trial was injected with 100  M. nipponense were obtained from the same farm as the adults and acclimated for two weeks in the laboratory conditions prior to the feeding trial. The basal diet was supplemented with copper sulphate  at 0, 10, 20, 30, 40, 80, and 160 mg Cu kg−1 diet, respectively. Procedure of diet preparation was similar to that described by Li et al. [ M. nipponense were included. The actual copper concentrations in the feeds were analyzed to be 2.8, 12.2, 20.9, 29.8, 43.1, 78.9, and 157.1 mg kg−1, respectively, by the flame atomic absorption photometry [The juvenilei et al. , and all−1, and total ammonia nitrogen <0.1 mg L−1. The Cu concentration in rearing water was 1.4–1.7 μg Cu L−1. At the end of the feeding trial, all prawns were counted and gained survival rate /). Prawns were fasted for 24 h before the hepatopancreas was collected. These samples were stored at −80°C until RNA extraction.Prawn juveniles (0.101 ± 0.001 g) were randomly placed in 21 of 300-L tanks with 30 prawns per tank in triplicate. Prawns were fed to apparent satiation twice daily (8:00 and 17:00 h) for 56 days. To maintain water quality, one-third of the tank water was exchanged daily. During the feeding period, water temperature was 27–30°C, dissolved oxygen >6.5 mg Lμg of total RNA was used for synthesis of first-strand cDNA by the PrimeScript™ RT-PCR Kit . Acquired cDNA was stored at −20°C for subsequent quantitative real-time PCR (qRT-PCR).Total RNA was isolated using RNA extraction kit  following manufacture protocol. In reverse transcription reaction, 3  M. nipponense [μL containing 5.0 μL 10x Ex Taq Buffer (Mg2+ Plus), 5.0 μL 10x UPM, 4.0 μL cDNA template, 4.0 μL dNTPs mix (2.5 mM each), 1.0 μL GSP (10 μM), 0.26 μL Ex Taq HS , and 30.74 μL of sterile deionized water. The PCR conditions were set as follows: 5 cycles of 94°C for 30 s and 72°C for 3 min, then 5 cycles of 94°C for 30 s and 70°C for 30 s, and 25 cycles of 94°C for 30 s, 68°C for 30 s, and 72°C for 3 min. All used primers were summarized in To get the full length of MnHc-1 cDNA sequence, SMART™ RACE cDNA Amplification Kit  was used to conduct the 3′-RACE and 5′-RACE. Gene-specific primers of MnHc-1 were designed based on the known fragments initially identified from the EST cDNA library ofpponense . The totThe PCR products were purified by the UNIQ-10 Gel Extraction Kit  and cloned into the pUCm−T vector . The transformed bacteria were identified, confirmed by blue/white screening, and validated by PCR. More than two recombinant plasmids were sequenced using BigDye Terminator v3.1 Cycle Sequencing Kit operating on an Automatic DNA Sequencer .http://www.ncbi.nlm.nih.gov/gorf/) was used to predict the gene's putative open reading frame. The BLAST algorithm at the National Center for Biotechnology Information (http://www.ncbi.nlm.nih.gov/BLAST/) was applied to search for sequence homology. SMART  was used to predict conserved motifs. Signal sequence was carried out using SignalP 4.1 program . The deduced amino acid sequence of MnHc-1 from M. nipponense and other invertebrate hemocyanin sequences acquired from NCBI database were aligned by software ClustalX. A neighbor-joining (NJ) phylogenetic relationship was established based on amino acid sequences of the hemocyanin using the MEGA 5.1 program (http://www.megasoftware.net/).The online ORF finder program  was used for statistical analysis. The results of relative mRNA expression in challenge test were analyzed by  M. nipponense was 2,163 bp (GenBank accession number JX456149.1), containing a 20 bp 5′-untranslated region, a 115 bp 3′-untranslated region with poly A tail, and a 2,028 bp open reading frame (ORF) (The full length of MnHc-1 cDNA from theme (ORF) . The ORF A. moluccensis gamma subunit, P. monodon, C. quadricarinatus, and E. sinensis, respectively. The MnHc-1 exhibited 64% identity with another subunit of hemocyanin in M. nipponense . A phylogenetic tree was constructed with the 24 full-length hemocyanin sequences from arthropod based on the neighbor-joining method ; the expression value is 20.32, which is 20 times higher or more compared with other tissues. The value is as low as 0 so that the gene is not expressed in muscle  (P < 0.05) and then started to significantly increase and reached the peak in 12 h (6.2 times) (P < 0.05), followed by a recovery to the initial level in 24 h and 48 h.−1 diet, the level of MnHc-1 mRNA in hepatopancreas of prawns significantly increased (1.9-fold to 5.8-fold) (P < 0.05) and was significantly higher compared to that fed 2.8–29.8 mg Cu kg−1 diet (P < 0.05), but differences in 2.8–29.8 groups were not significant (P > 0.05).Survival rate (70–81%) of prawns was not affected by the dietary copper levels. As shown in  M. nipponense (MnHc-1). MnHc-1 was a polypeptide of 675 amino acids with a 21-amino acid putative signal peptide. The signal peptide ends Ala-X-Ala motif, which is a frequent accordance prior to the cleavage site of signal peptides, suggesting that a cleavage site is located at the 21-22 amino acids [Hemocyanin is an extracellular, multisubunit protein in crustacean , 33. Thono acids , 35. Strno acids , 14, 15.no acids . The phyno acids . H. americanus [ Fenneropenaeus chinensis [ E. sinensis [ M. nipponense (MnHc-2) expressed in the muscle [The present study showed that the highest level of MnHc-1 mRNA expression occurred in the hepatopancreas. The result is the same as the findings in other crustaceans, such asericanus , Fennerohinensis , and E. sinensis , and consinensis . MnHc-1 e muscle . We alsoe muscle . The dis Vibrio alginolyticus, Vibrio harveyi, A. hydrophila, and Staphylococcus aureus in L. vannamei serum was hemocyanin, suggesting that hemocyanin possesses antibacterial functions. The C-terminus of L. vannamei hemocyanin is possibly related to the immunity in shrimp to different pathogens [ A. hydrophila are a common species of the Aeromonas genus in water and water habitants [ A. hydrophila in fish and prawn including M. nipponense has been one of the major diseases under farming conditions [ E. sinensis was significantly upregulated by A. hydrophila infection. In our study, temporal and spatial expressions of MnHc-1 in the hemocytes of prawn infected with A. hydrophila showed a clear time-dependent pattern. The level of MnHc-1 mRNA expression significantly decreased at 3 h after injection, then started to significantly increase after 6 h and 12 h, and then reached the peak at 12 h, implying that the hemocyanin is involved in the antibacterial defense of prawn. In another study, the level of MnHc-2 mRNA expression in prawn significantly increased over time and peaked at 3 h after the A. hydrophila challenge [ A. hydrophila injection. Lei et al. [ P. japonicus that PjHcL transcriptional upregulation occurred after 4 h of injection of the active WSSV. It is possible that PjHcL may be triggered by the fast expressed proteins in virus. It is also likely that MnHc-1 may be induced by fast expressed protein in bacteria, but the detailed defensing mechanism of hemocyanin against bacterial infection needs further study.Hemocyanin is an important multifunctional protein in mollusks and arthropods. Besides its role as an oxygen carrier, its immune functions including antibacterial activities, agglutination property, and PO activity have become hot topics of immunological research , 4, 5, 7athogens . Gram-neabitants . The infnditions , 41. Sunnditions  showed thallenge . Clearly E. sinensis, expression of hemocyanin mRNA was affected by the level of dietary copper [−1 diet was significantly higher compared to that fed 2.8–29.8 mg Cu kg−1. The increase of dietary Cu concentration also increased the Cu content in hepatopancreas, especially in the 43.1–157.1 mg Cu kg−1 groups. Hemocyanin synthesis mainly occurs in the hepatopancreas [−1 diet) can increase hemocyanin mRNA expression in the hepatopancreas and hemocytes of crab [The expression of hemocyanin subunits varies with environmental or nutritional changes , 43. Copy copper . Our prepancreas . Therefo M. nipponense. Our results suggest that MnHc-1 may play a critical role in antibacterial defense in prawn. Accumulation of high copper levels in hepatopancreas of prawn triggers MnHc-1 gene expression. These results provide the foundation for further studies in biological function and regulation of hemocyanin in crustacean.In conclusion, we cloned the hemocyanin subunit gene (MnHc-1) from the hepatopancreas of


The Seebeck coefficient increased a great deal with increasing x, due to decreasing carrier concentration, and the total thermal conductivity reduced mainly because of declining κe. Finally, a peak thermoelectric figure of merit, ZT, was about 0.46 for NbCo1.3Sb at 973 K. This enhancement was mainly attributed to the reduction of electric thermal conductivity and the increase of Seebeck coefficient. The excess Co had effects on the carrier concentration, deformation potential Edef and DOS effective mass m*. Adding an excessive amount of Co leads to a very high Edef, which was detrimental for transport characteristics.NbCoSb with nominal 19 valence electrons, and is supposed to be metallic, has recently been reported to also exhibit the thermoelectric properties of a heavily doped n-type semiconductor. In this study, we prepared Co-rich NbCo ZT = S2σT/(κe+ κL), where S, σ, T, κe, and κL are the Seebeck coefficient, electrical conductivity, absolute temperature, and electronic and lattice components of the thermal conductivity, respectively. It is hard to achieve a high ZT by simply improving one of these parameters, because S, σ, κe are strongly coupled with each other [Thermoelectric (TE) materials can directly convert waste heat into electricity in an environmentally-friendly way and have received a great deal of attention owing to global energy sources and environmental challenges . The conch other . κL, such as introducing secondary phase [2σS) by optimizing carrier concentration [Usually, an ideal thermoelectric material should have a glass-like heat conduction of phonons and crystal-like electrical conduction of charge carriers. This strategy is referred to as “phonon-glass electron-crystal”, and was first introduced by Slack . To imprry phase , nanoscary phase  and poinry phase . To enhantration  or energntration .ZT of about 1.5 at 1200 K has been realized in p-type NbFeSb alloys [ZT of ~0.4 at 973 K [Recently, particular attention has been paid to half-Heusler (HH) compounds, due to their high-temperature stability, good mechanical robustness, and good TE performance, which are of paramount importance for practical applications. Typical semiconducting HH compounds with 18 valence electrons per unit cell, ZrCoSb, ZrNiSn, TiNiSn, NbFeSb, and their alloys have been intensively investigated as promising medium–high temperature TE materials ,14,15,16b alloys ,18. Surpat 973 K . The expZT of ZrNi1.1Sn was approx. 40% higher compared to ZrNiSn, due to the secondary phase in the matrix introduced by excess Ni. It provided a great basis for phonon-scattering centers, leading to an improved Seebeck coefficient and a reduced thermal conductivity [x1+Sb  were prepared through arc melting, following by ball milling and hot pressing. The evaluation and comparison of phase composition and TE properties of NbCox1+Sb were performed. Additionally, the electrical transport characteristics of Co-rich NbCoSb materials were also analyzed using the single parabolic band (SPB) model.From our previous work, we know that NbCoSb has a comparatively low Seebeck coefficient and a higher thermal conductivity. According to Chai’s report ,21, ZT ouctivity ,21. Inspx1+Sb . For all samples, 5 at% more Sb was added in order to compensate the loss of Sb during arc melting. In order to get homogeneous ingots, metallic raw materials were arc-melted five times reversely under argon protection. The obtained ingots were ball milled  for 7 hours under argon protection. Thereafter, the ball-milled powders were loaded into a graphite die and sintered by direct current-induced hot pressing at 1273 K for 2 min under 77 MPa to get pellet samples. The thickness of the as-hot-pressed samples was about 2 mm.A series of Co-rich NbCoSb samples were prepared by arc melting followed by ball milling and hot pressing. High purity niobium slugs , cobalt pieces , and antimony rods  were weighed according to the composition of NbCoκ = pdDC was calculated using the measured density (d) by Archimedes method, thermal diffusivity (D) by laser flash method , and specific heat capacity (pC) by differential scanning calorimetry . Bar-shaped samples of about 2 mm × 2 mm × 10 mm were used for measuring the electrical conductivity and Seebeck coefficient simultaneously from 300 to 973 K . The Hall coefficient (RH) was measured at room temperature by a commercial system  using a four-probe configuration and the Van der Pauw method, with a magnetic field of 3.0 T and an electrical current of 8 mA. The carrier concentration (n) was calculated by n = 1/(eRH), and the carrier mobility (μ) was estimated by σ = neμ. The properties of the samples were isotropic, due to their small grain size and random crystal orientation.The thermal conductivity 1.3Sb was studied via SEM  to show the particle size.The phase structure of the samples was observed using XRD on a PANalytical X’Pert Pro diffractometer  and Cu Kα radiation. The fresh section of the hot-pressed sample NbCox1+Sb  which can be classified as cubic MgAgAs-type crystal structure . The dominant phase of all samples was the half–Heusler phase of NbCoSb, while a small number of secondary phases such as Nb3Sb, Nb1.08Co1.92, and NbCo2 were observed following the increasing content of excess Co, respectively. Based on the binary phase diagram of Nb and Sb, Nb3Sb may be formed, due to the peritectic reaction during the cooling process (x = 0), as shown in x = 0.2, 0.3), the secondary phases Nb3Sb disappeared. However, another secondary phase Nb1.08Co1.92 was introduced in a small amount. A new secondary phase NbCo2 was observed for the Co-rich sample NbCo1.4Sb, with increased content of excess Co. Even more NbCo2 appeared for NbCo1.5Sb. These phenomena indicate a scale of Co solubility in the structure. However, when x exceeded 0.3, it precipitated out the other second phase of NbCo2.x1+Sb samples. The lattice parameter increased with increasing x, mainly due to the solid solution of excess Co, leading to lattice expansion of HH phase. The theoretical density was calculated by dcal = ∑niMi/(a3AN), where ni is the number of atoms of Nb, Co and Sb per unit cell, Mi the corresponding atomic mass of each element, a the lattice parameter, and AN the Avogadro constant (6.023 × 1023 mol−1). The relative density was more than 90% for x = 0, 0.2, 0.3 and approx. 85–86% for x = 0.4 and 0.5. The low relative density of the NbCo1.4Sb and NbCo1.5Sb samples may be due to inaccurate estimation of the theoretical density. The additional Co did not fully enter the lattice, but form the second phase, while we ignored this in our calculations.1.3Sb, indicating a high relative density. Moreover, the grainsize of the sample was approx. several hundreds of nanometers.As shown in x1+Sb. The electrical conductivity of NbCox1+Sb decreased with increasing temperature for x = 0, 0.2, 0.3, indicating the characteristics of metal  at room temperature was calculated based on Equations (1)–(6) using a SPB model. The results are shown in nF(η) is the nth order Fermi integral, η the reduced Fermi energy, e the electron charge, r the scattering factor, Bk the Boltzmann’s constant, h the Plank’s constant, Hr the Hall factor, and x the variable of integration. r = −1/2 was used for the acoustic phonon scattering mechanism. Combining the experimental carrier concentrations and Seebeck coefficients, a density of states (DOS) effective mass m* ~8.5 me was derived for Co-rich samples NbCox1+Sb. Therefore, the improvements of Seebeck coefficients for Co-rich samples could be explained by their relatively high DOS effective mass m* and reduced carrier concentration.of metal a. Howeventration . The powat 973 K c. The pox. The mobility of the Co-rich samples was largely decreased for x = 0.4 and 0.5, which may be due to their enhanced effective mass and existence of secondary phases. defE specifically characterizes the scattering effects of lattice vibration on carriers, namely the coupling effects between phonons and electrons. defE can be roughly estimated by Equation (7) when acoustic phonon scattering is dominant [d is the density (approx. 8.8 g cm−3), lv the longitudinal velocity of sound (approx. 5488 m s−1), µH the drift mobility (µH = µ/rH), and Nv the number of degenerate carrier pockets. The deformation potential defE of NbCo1.5Sb was found to be 41.38 eV (0.8Sn0.2 (approx. 21 eV) [The deformation potential dominant ,25.(7)E41.38 eV , a relat. 21 eV) .x1+Sb. Obviously, diffusivity showed a negative linear relationship with temperature and slightly decreased after adding excess Co except for x = 0.5 –(3)—combines the measured Seebeck coefficients. The calculated Lorenz constant was approx. 2.0 × 10−8 W Ω K−2 for NbCo1.3Sb at room temperature. κL could be calculated by subtraction of κe from κ. As shown in κL increased slightly for Co-rich samples, while κe dropped steadily over the whole measured temperature range. Here, the reduction of κe can be primarily ascribed to both the reduction of L and σ. The obvious decrease of κe happened due to reduced carrier concentration, contributing to a reduced Lorenz constant. Typically, κe at room temperature decreased more than 70% from 2.34 W m−1 K−1 for NbCoSb to 0.19 W m−1 K−1 for NbCo1.5Sb. Moreover, the slight increase of κL may be due to weakened electron-phonon scattering, which became weaker for Co-rich samples [1.08Co1.92 and NbCo2, or other possible phonon scattering.  x = 0.5 a. As sho samples . It mighminκ) of NbCoSb using Cahill’s method, assuming the phonon’s mean free path is half of the phonon’s wavelength [V, tν, and lν are average volume per atom, transversal and longitudinal wave velocity , respectively. In general, κ was found to be larger than the estimated minκ ≈ 0.9 W m−1 K−1. It seems that there is a large potential to improve the ZT values by further reducing the lattice thermal conductivity of NbCoSb based TE materials, definitely, further efforts are required to explore new useful ways to enhance the thermoelectric performance.Furthermore, we estimated the minimum lattice thermal conductivity (velength :(8)κminZT was calculated as a function of temperature as shown in ZT of 0.46 was obtained at 973 K for sample NbCo1.3Sb. A slight increase of ZT should contribute to decreased thermal conductivity and a moderate PF.Combining all transport properties, the figure of merit x1+Sb  were synthesized and characterized. A small change in lattice parameter indicated a certain degree of Co solution into half-Heusler matrix. Adding excess Co eliminated the secondary phase of Nb3Sb, while other impurity phases such as Nb1.08Co1.92 and NbCo2 appeared with more excess Co. With increasing x, the Seebeck coefficient increased a lot due to decreasing carrier concentration; total thermal conductivity decreased mainly because of declining κe. Adding an excessive amount of Co leads to a very high deformation potential Edef, such as NbCo1.5Sb (41.38 eV), which is detrimental for transport characteristics. Finally, a maximum ZT value of approx. 0.46 for NbCo1.3Sb was obtained at 973 K.In summary, Co-rich samples NbCo


Amphora sp. was isolated from the Sfax Solar Saltern and cultivated under hypersaline conditions. It contains moderate rates of proteins, lipids, sugars, and minerals and a prominent content of bioactive compounds: polyphenols, chlorophyll a, carotenoids, and fatty acids. The analysis of fatty acids with GC/MS showed that the C16 series accounted for about 75% of Amphora sp. lipids. Saturated fatty acids whose palmitic acid was the most important (27.41%) represented 41.31%. Amphora sp. was found to be rich in monounsaturated fatty acids with dominance of palmitoleic acid. It also contains a significant percentage of polyunsaturated fatty acids with a high amount of eicosapentaenoic acid (2.36%). Among the various solvents used, ethanol at 80% extracted the highest amounts of phenols and flavonoids that were 38.27 mg gallic acid equivalent and 17.69 mg catechin equivalent g−1 of dried extract, respectively. Using various in vitro assays including DPPH and ABTS radicals methods, reducing power assay, and β-carotene bleaching assay, the 80% ethanolic extract showed high antioxidant activity. A strong antibacterial activity was checked against Gram-positive bacteria (Staphylococcus aureus and Micrococcus luteus) and Gram-negative bacteria . These results are in favor of Amphora sp. valorization in aquaculture and food and pharmaceutical industries. Microalgae are considered as an important source of bioactive compounds with a wide range of applications and commercial use. They produce proteins, lipids, vitamins, pigments, and other molecules exploited for health, for food and feed additives, and for energy production. Due to their wide diversity and high storage capacity for lipids, particularly eicosapentaenoic acid (EPA) and amino acids, microalgae have pharmaceutical and cosmetic applications , 2. FurtAmong the large variety of microalgae, Cyanophyceae, Chlorophyceae, Bacillariophyceae (diatoms), and Chrysophyceae are the most studied for biodiesel production . It is eUnlike the universal macromolecules of primary metabolism , which are commonly present in all organisms, secondary metabolites have a far more limited distribution in nature. They are not necessarily produced under all conditions and can only be found in specific organisms or a group of organisms. The organism can produce these compounds either to protect itself within its own living ecosystem or to play a basic role in its everyday existence. This reveals bioactive molecules in unrelated biological systems . The med Amphora sp. (Bacillariophyceae) was cultivated in a hypersaline medium and the physicochemical properties, fatty acids profile, bioactive compounds, and some biological activities were determined.In the present work,Amphora sp. is a Bacillariophyceae that was isolated via micromanipulation and serial dilution from a water sample collected in the pond C4-1 of the Sfax Solar Saltern with average salinity of 107 p.s.u. . The Sfax Solar Saltern is located in the central eastern cost of Tunisia, at about 34°39′N and 10°42′E. The ponds are shallow (20–70 cm depth) and have various salinities ranging from 40 to 400 p.s.u. . The mixture was incubated at 50°C for 20 min, and then 1.25 mL of 10% (w/v) trichloroacetic acid was added to the mixture which was then centrifuged at 3500 ×g for 10 min. The supernatant (1.25 mL) was mixed with 1.25 mL distilled water and 0.25 mL of 0.1% FeCl3 (w/v). The absorbance was measured at 700 nm. Ascorbic acid was used as a reference. The increase of the absorbance in the reaction indicates the increase of the iron reduction. The values presented are the mean of triplicate analysis.The reducing power of the extract was determined according to the method described by Yildirim et al. . Brieflyβ-carotene was assessed as described by Koleva et al. [β-carotene/linoleic acid mixture was prepared by dissolving 0.5 mg of β-carotene in 1 mL of chloroform, 25 µL of linoleic acid, and 200 µL of Tween 80. The chloroform was completely evaporated under vacuum in a rotary evaporator at 40°C, and then 100 mL of distilled water was added and the resulting mixture was vigorously stirred. The emulsion obtained was freshly prepared before each experiment. Aliquots (2.5 mL) of the β-carotene/linoleic acid emulsion were transferred to test tubes containing 0.1 mL of extract at different concentrations (0.1–0.5 mg mL−1). Following incubation for 2 h at 50°C, the absorbance of each test was measured at 470 nm. Vitamin C was used as a positive standard. The control tube contained no sample. Antioxidant activity in β-carotene bleaching model expressed as percentage was calculated with the following equation:A0 and A′0 are the absorbance of the sample and the control, respectively, measured at time zero and At and A′t are the absorbance of the sample and the control, respectively, measured after 2 hours. Tests were carried out in triplicate.The ability of the extract to prevent the bleaching of a et al. . A stockBacillus cereus (ATCC11778), Bacillus subtilis (JN934392), Micrococcus luteus (ATCC4698), and Staphylococcus aureus (ATCC6538)) and Gram-negative (Klebsiella pneumoniae (ATCC13883), Salmonella enterica (ATCC43972)) bacteria. The antimicrobial activity of 80% ethanolic extract was evaluated using agar well diffusion method. The wells were then filled with 60 µL of the extract at 20 mg/mL in 5% DMSO and 5% DMSO was used as negative control. Besides, plates were incubated for 24 hours at 37 ± 1°C for bacterial strains. Then, the diameters of inhibition zones (IZD) were measured.The operative pathogens were Gram-positive  [Some physicochemical characteristics ofTM 20023  and 5.9%eaformis . However Amphora , and it 6.9% DW) . A study6.9% DW) . Amphora sp. (27.62%) was lower than that found for other strains of Amphora; 54% for CTM 20023 [ Amphora sp. [ Amphora coffeaformis [The proteins level forTM 20023  and 30–4hora sp. . Indeed,hora sp.  and higheaformis . Amphora sp. was 12.60% DW, which is consistent with that of some microalgae (5–23% DW) [ Amphora coffeaformis and some diatoms contain a high amount of sugars in the range of 13.5–16.4% [The total sugars content of–23% DW) . Neverth.5–16.4% . Amphora sp. (37.78% DW), it is in line with that found for another Amphora strain (30% DW) cultivated at salinity ranging between 15 and 35 p.s.u. [ Amphora sp. has moderate amounts of sodium, potassium, calcium, and magnesium.With respect to the ash content of5 p.s.u. . Ash con5 p.s.u. . Amphora Amphora sp. was found to be rich in chlorophyll, mainly chlorophyll a, and carotenoids. In fact, its chlorophyll content reached almost 5% of the dry matter, which is in agreement with other research works [β-carotene and xanthophylls, thus indicating a powerful antioxidant activity [Moreover,ch works . Furtheractivity . Amphora sp. was composed of saturated, monounsaturated, and polyunsaturated fatty acids , respectively  and contains an important percentage of PUFA (6.03%). High levels of palmitoleic acid and other bioactive fatty acids were also detected in the fusiform morphotype of the Bacillariophyceae Phaeodactylum tricornutum [In salinity of 100 p.s.u., on F/2 medium, the fatty acids profile ofectively . The C16iodiesel . Amphoracornutum . Eicosapcornutum  and canccornutum . Amphora sp. extracts are summarized in −1 extract and 17.69 ± 0.70 mg CE g−1 extract, respectively, compared to the extracts obtained by ethyl acetate, hexane, and water. The phenolic content of ethanol extracts of some Moroccan marine microalgae was found to range from 8.2 to 32 mg GAE g−1 extract [ Nannochloropsis gaditana with 32 mg GAE g−1 extract. The extracts of Dunaliella sp., Phaeodactylum tricornutum, and Navicula sp. also contained high phenolic content of more than 15 mg GAE g−1 extract [The total phenols and flavonoids contents of extract . The aut extract . Amphora sp. was 3.22 mg CE g−1 DW, which is sixfold higher than that found for Amphora CTM 20023 [The flavonoid content in the 80% ethanolic extract ofTM 20023 . Amphora sp. 80% ethanolic extract may be due to the culture conditions under the high salinity of 100 p.s.u. and the extraction conditions.The high levels of polyphenols and flavonoids in the in vitro assays: the DPPH and ABTS radical scavenging capacities, the ferric reducing antioxidant power, and the β-carotene bleaching inhibition tests, using different concentrations of the extract. The inhibition percentages of the radicals DPPH and ABTS by the extract and standards (Trolox and BHT) were found to be concentration-dependent (0.065 to 1 mg mL−1 for DPPH test and 0.5 to 4 mg mL−1 for ABTS test) as shown in 50 are presented in 50 indicates a high antioxidant activity. Amphora sp. 80% ethanolic extract exhibited high antiradical power with an IC50 value of 0.23 ± 0.07 mg mL−1 for DPPH scavenging activity and an IC50 value of 2.61 ± 0.64 mg mL−1 for ABTS radical scavenging capacity. Some authors have found a strong correlation between the phenolic contents and the antioxidant activity [Due to its highest content of phenols and flavonoids, the 80% ethanolic extract was used to check antioxidant activity through fouractivity .The FRAP assay is based on the ability of phenols to reduce yellow ferric tripyridyl triazine complex (Fe(III)-TPTZ) to blue ferrous complex (Fe(II)-TPTZ) by the action of electron-donating antioxidants .  Figure Amphora coffeaeformis, Navicula sp., and Achnanthes longipes, resp.) [50 value for ABTS scavenging capacity of Amphora sp. extract was 2.6 mg mL−1. Many research studies [50 ABTS scavenging capacities for two brown algae of 5.29 mg mL−1 for Sargassum binderi and 15.2 mg mL−1 for Turbinaria conoides.The effect of antioxidants on DPPH free radical scavenging was considered to be emanating from their hydrogen-donating ability. In this investigation, the 80% ethanol extract was proven to exhibit a strong effect of DPPH free radical scavenging. In fact, it reached 74.34% of inhibition at 1 mg of extract, which is considered higher than the methanol extract of some Bacillariophyceae tested at 2 mg of the extract  were measured at concentrations of extract between 0.1 and 0.5 mg mL−1, showing its dependency on concentration , with about twice the IC50 value of vitamin C used as the standard. The absorbance at 700 nm reached a value of 0.78 at a concentration of 0.5 mg mL−1 of the extract, which is considered high, indicating a good antioxidant activity. Therefore, Amphora sp. extract was confirmed to have a strong antioxidant capacity. The multiple regression analysis has proven that both carotenoid and phenolic contents significantly contribute to the explanation of the variation in FRAP and TEAC activity of the extracts. The regression coefficients indicate that phenols and carotenoids are of similar importance in explaining variation in the antioxidant activity [The ntration . The IC5ing test  was stroactivity . Amphora sp. 80% ethanolic extract was screened against six bacteria and their potency was qualitatively and quantitatively analyzed by the diameters of the inhibition zones (IZD) as summarized in  Amphora sp., at a concentration of 20 mg mL−1, was active against Gram-positive bacteria, Micrococcus luteus and Staphylococcus aureus, and inactive against both strains of Bacillus (cereus and subtilis). This extract was active against Gram-negative bacteria tested: Klebsiella pneumonia and Salmonella enterica. The extract exhibited strong activity against Micrococcus luteus, Staphylococcus aureus, and Klebsiella pneumoniae. The IZD for Klebsiella pneumoniae and Staphylococcus aureus of 16 mm and 20 mm, respectively, were stronger than those found for the ethanolic extract of a strain of Amphora sp. cultured in standard Walne's medium (10 mm and 10.5 mm), respectively [In the present study, the antibacterial activity ofectively . This diectively . Chaetoceros muelleri was related to its lipids composition [ Phaeodactylum tricornutum [ Skeletonema costatum [ Haematococcus pluvialis [ Dunaliella salina [The extract used in this study was found to be rich in a variety of compounds: lipids and fatty acids, carbohydrates, polysaccharides, polyphenols, flavonoids, pigments, and carotenoids, which can be indicative for having antibacterial activity as mentioned for several microalgae . The antposition . Fatty acornutum , 53 and costatum  and otheluvialis  and Dunaa salina .Microalgae extracts should be tested as additives for food and feed formulations, in an attempt to replace the currently used antimicrobial compounds of synthetic origins, including subtherapeutical doses of antibiotics employed as a prophylactic measure in animal breeding .Amphora sp., a brown microalga cultivated in high salinity medium of 100 p.s.u., was confirmed to contain a moderate percentage of proteins, lipids, sugars, and minerals and important levels of polyphenols, flavonoids, chlorophyll, carotenoids, and bioactive fatty acids as EPA. Besides, this microalga has important antioxidant and antibacterial activities. Prominent contents of bioactive compounds in this microalga are in favor of its possible valorization in food additives and pharmaceutical or cosmetic products. Moreover, its richness in saturated fatty acids allows its eventual use for biodiesel production.


Levosimendan exerted favorable effects on the initial outcome in the treatment of ventricular fibrillation cardiac arrest. This study investigated the efficacy of levosimendan in the treatment of asphyxia-induced cardiac arrest in rats.Animals underwent asphyxial cardiac arrest/cardiopulmonary resuscitation, randomized to three treatment groups: epinephrine (10 μg/kg) supplemented with levosimendan ; epinephrine only , or levosimendan only . The resuscitation success rate, wet-to-dry ratio of lung, and rate of alveolar and blood gas analysis were recorded.P < 0.001); among them, 10, 4, and 2 rats survived at the end of a 60-min observation period from each group, respectively (P = 0.001). The coronary perfusion pressure in the EL group was higher than that of either the E or L group (P < 0.05). The lung wet-to-dry weight ratio and rate of damaged alveoli were lower in the EL group than the E group (P < 0.05).10 rats in the EL group, 8 in the E group, and 2 in the L group showed an initial return of spontaneous circulation  contains supplementary material, which is available to authorized users. Cardiac arrest (CA) is the most devastating adverse cardiac event, and it causes approximately 325,000 deaths each year in the United States . Asphyxi2+-saturated troponin C of the myocardial thin filament , and the epinephrine-only treatment group (E), epinephrine (10 μg/kg in 2.5 ml of volume) administered with saline 0.9% [bolus during CPR and infusion for 1 h ] or levosimendan-only treatment group (L), saline 0.9%  administered with levosimendan [bolus 12 μg/kg in 1.2 ml of volume and infusion for 1 h (0.3 μg/kg/min in 0.03 ml of volume)].2,1.0; respiratory rate, 75–80 breaths/min; inspiratory/expiratory ratio, 2:3; HX-300; TME Technology Co., Ltd., Chengdu, China) [All rats were fasted for 12 h before the experiment began and were given access to water ad libitum. They were anesthetized with an intraperitoneal injection of urethane . A tracheal intubation was performed via tracheotomy and rats were connected to the rodent volume-controlled ventilator  , 16. The, China) . Arteria, China) .The asphyxia-induced CA model was quoted from the experiment performed by McCaul et al. . After a2 1.0, VT 8 ml/kg, rate 75–80 min−1), beginning external chest compressions (approximately 300 min−1 at a depth of 1 cm), and through intravenous administration of medication. Effective resuscitation (ROSC) was defined as a native RPP more than 20% of baseline value for 1 min [Resuscitation began 1 min after onset of CA and was established by starting the ventilator . Finally, arterial blood gas analyses were performed.The animals were sacrificed after CPR. The lobe of the left lung was weighed and dried, and we calculated the lung wet-to-dry weight ratio. We fixed the middle lobe of the right lungin10% formalin fixative, created paraffin-embedded slices, and used hematoxylin and eosin stain for observation . Fifty vBased on our pre-test, we used the Power Sample Size (PASS11.0) software program for power analysis and compared the survival rates among the experimental groups. In our preliminary study, 18 rats were used, with 6 rats in each group. There were 6, 1, and 1 rat in groups EL, E, and L that survived to 60 min, respectively. The survival rate was 100, 16.7, and 16.7%, respectively. We set the power at 0.8; the significance criterion was 0.017. As a result, we needed 8 rats per group to achieve statistical significance. To account for potential attrition, we enrolled 10 rats per group.P value < 0.05 was considered significant. Fisher probabilities in a 3 × 2 table were used for analyzing success resuscitation rates and survival rates, and P value < 0.017 was considered significant in post hoc multiple comparisons.All data were performed using SPSS for 17.0 for Windows. All data generated or analysed during this study are included in this published article . Further comparisons in the first 3 min, e.g., the CPP and MAP, but not the RPP or HR, in the EL group were higher than that in the E or L group (P < 0.05).Hemodynamic values such as CPP, MAP, HR, and RPP for the three groups are presented in Fig. P = 0.005). The ratio in the EL group was significantly lower than that of the E group . The rate of damaged alveoli in the EL group was significantly lower than that of the E group (P < 0.001).In the EL group, there was no evidence of alveolar structural damage, edema, or hemorrhage. In the E group, there were numerous erythrocytes in the alveolar field, accompanied by altered alveolar structures. In the L group, some alveoli could be identified, and there were numerous erythrocytes observed in the injured alveoli Fig. . The EL,P < 0.05). Specifically, the PH, PaO2, HCO3-, and base excess (BE) values of the EL group were significantly higher than that of the E group . In addition, the PH values and BE in the EL group were significantly higher than that of the L group . The blood lactate value in the EL group was significantly lower than that of the E or L group (EL vs. E, P = 0.018, EL vs. L, P = 0.001; Table Arterial blood gas parameters at 60 min are shown in Table In our study of resuscitation on asphyxia-induced CA, we demonstrated that CPP significantly improves when rats received epinephrine therapy supplemented with levosimendan. The EL group also exhibited a significant reduction in lung injury and acidosis, excellent oxygen partial pressure, and an improved survival rate compared with rats that received epinephrine or levosimendan alone.CPP, calculated as aortic pressure minus right atrial pressure during the diastolic phase of CPR, is strongly associated with resuscitation outcomes , 21. Fri2+ inflow, reduced the sarcoplasmic reticulum uptake of release of Ca2+ and hindered the binding between troponin and Ca2+. Levosimendan is a selective systolic calcium sensitizer mainly used clinically for the treatment of acute heart failure. When sudden CA occurs, synthesis of ATP is blocked and myocardial cell cannot pump redundant Ca2+ to the extracellular space, thereby inhibiting excitation contraction coupling. Myocardial cytoclasis then occurs due to an overload of calcium. If CPR is implemented immediately, myocardial perfusion would maintain calcium ion at 25 to 30% of normal levels. Epinephrine is added at the same time to constrict the blood vessels to enhance CPP. In addition, levosimendan has a direct vasodilatory effect on coronary arteries and enhances coronary blood flow [Myocardium contraction weakened when heart failure, correlated with the decrease in extracellular Caood flow , 24. In Krishnamoorthy et al.  reportedMany α-agonist agents have been shown to increase CPP. However, no placebo-controlled study has shown that the use of any vasoconstrictor during CA can increase survival rates . Kelm etIt should be recognized that this study has limitations. Our study focused exclusively on short-term survival results and does not address neurological outcomes and long-term survival outcomes, which are the end point of cardiopulmonary cerebral resuscitation in clinical practice. We will explore these parameters in future experiments.As a calcium-sensitizer agent, levosimendan improved initial resuscitation outcomes in asphyxia-induced CA when administered with epinephrine during CPR. These effects may contribute to increased coronary perfusion flow and reduced lung injury and acidosis. Our study has suggested that levosimendan can be selected as a promising alternative supplement agent to epinephrine during CPR for asphyxia-induced CA.


Ambulance services and stroke alerts reduce the time from stroke onset to acute stroke diagnosis. We describe the use of stroke alerts and ambulance services in different hospitals and patient groups and their relationship with reperfusion therapy.This nationwide study included 49,907 patients admitted with acute stroke who were registered in The Swedish Stroke Register (Riksstroke) in 2011–2012.n = 9), 0.5% to 38.7% in specialized nonuniversity hospitals (n = 22), and 4.2% to 40.3% in community hospitals (n = 41). Younger age, atrial fibrillation (AF), living in an institution, reduced consciousness upon admission, and hemorrhagic stroke were factors associated with a higher probability of stroke alerts. Living alone, primary school education, non‐European origin, previous stroke, diabetes, smoking, and dependency in activities of daily living (ADL) were associated with a lower probability of stroke alert. The proportion of patients arriving at the hospital by ambulance varied from 60.3% to 94.5%. Older age, living alone, primary school education, being born in a European country, previous stroke, AF, dependency in ADL, living in an institution, reduced consciousness upon admission, and hemorrhagic stroke were associated with ambulance services. Hospital stroke alert frequencies correlated strongly with reperfusion rates (r = .75).The proportions of patients admitted as stroke alerts out of all acute stroke admissions varied from 12.2% to 45.7% in university hospitals (Acute stroke alerts have a significant potential to improve stroke reperfusion rates. Prehospital stroke management varies conspicuously between hospitals and patient groups, and the elderly and patients living alone have a markedly reduced likelihood of stroke alerts. All hospitals admitting acute stroke patients in Sweden participate , and Riksstroke has an estimated coverage of 94% of all acute stroke patients treated in Swedish hospitals (Riksstroke). Riksstroke includes information on living conditions and comorbidities prior to stroke, acute stroke treatment, and secondary prevention, and patient‐reported outcome is followed up after 3 and 12 months. Details on what information is collected are available at the Riksstroke website The main analysis in this study included all 49,907 stroke events  in patients ≥18 years that were registered in Riksstroke in 2011–2012. The descriptive analysis of time trends in stroke alert frequencies included 200,133 stroke events registered from 2005 to 2012.2.2All Swedish hospitals have stroke alert protocols to quickly and accurately identify and triage stroke patients. SOS Alarm, the service coordinating ambulance calls in Sweden, is classifying stroke as a level 1 priority, irrespective of the time from onset , comorbidities , institutional living, smoking status, level of consciousness at admission, stroke subtype, and outcome . To adjust for individual hospital variation , hospital was included and modeled as a fixed effect  in the multiple regression. Patients with other missing variables were omitted from the multiple regression analysis. A difference with a 33.1p < .001). In the two most recent years, 2011 and 2012, the frequency was 27.5%  in university hospitals, 21.7% (range: 0.5%–38.7%) in specialized nonuniversity hospitals, and 18.3% (range: 4.2%–40.3%) in community hospitals. Compared with community hospitals, stroke alert frequencies were higher in specialized nonuniversity hospitals  and in university hospitals  after adjustment for patient‐level factors  in university hospitals, 72.8% (range: 66.9%–80.3%) in specialized nonuniversity hospitals, and 72.7% (range: 60.3%–94.5%) in community hospitals.p < .001).Older age, living alone, primary school education, European origin, previous stroke, AF, dependency in ADL, living in an institution, being drowsy or unconscious at admission, and hemorrhagic stroke were factors associated with use of ambulance services in the multiple regression Table . Patient3.3In total, 15,904 (31.9%) of the patients were admitted to a hospital within 3 hr of the onset of symptoms, 3,838 (7.7%) within 3–4.5 hr, 14,062 (28.2%) within 4.5–24 hr, and 7,747 (15.5%) later than 24 hr from onset. OAT was unknown for 8,356 (16.7%) of the patients.In the multiple logistic regression, adjusting for other patient factors, an OAT within 3 hr was associated with younger age, secondary or university education, being married or cohabiting, having AF, not having diabetes, nonsmoking, living in an institution, being independent in ADL, being drowsy or unconscious on admission, and suffering from hemorrhagic stroke  received reperfusion therapy . The reperfusion rate was 15.0%  in university hospitals, 10.7% (range: 6.1%–15.7%) in specialized nonuniversity hospitals, and 10.5% (range: 2.6%–22.3%) in community hospitals performing thrombolysis.r = .75, p < .001, Figure r = −.02, p = .84).There was a strong correlation between hospital stroke alert frequency and reperfusion rate . The use of ambulance services follows a partly different pattern with similar average proportions in university and nonuniversity hospitals, and a predominance of elderly patients with a low level of education. Common to ambulance services and stroke alerts is that they are used more infrequently with immigrants born outside Europe and, as expected, more often with patients with severe stroke.Most previous studies on stroke alerts have been performed in single centers  and because of more uncertainties about onset of stroke symptoms. As a result, fewer patients living alone were admitted to hospital within 3 hr of onset of stroke symptoms (present results), and 44% fewer received thrombolytic treatment (Stecksen et al., 4.1Strength of this study is that it is nationwide and covers all hospitals admitting acute stroke patients in the country and that it has high coverage of all patients admitted to hospital for acute stroke in the country. This study was based on individual‐level socioeconomic information, like the GWTG‐Stroke study (Lin et al., A limitation of this register‐based study is that it was not possible to distinguish between stroke alerts elicited by ambulance services (also called prehospital triage [Prabhakaran et al., Results from observational studies like the present one raise the question of possible residual confounding variables that might have influenced some or all of the findings. There are, for instance, many aspects of living conditions that might affect the use of stroke alerts and ambulance services, for example, distance to hospital, that might affect the use of stroke alerts and ambulance services, but these have not been measured. Riksstroke did not include a stroke scale during the study period, and hence we could not specify which stroke signs influenced stroke alerts. There was no information on stroke size or location. Level of consciousness at hospital arrival was used as a surrogate for stroke severity.5Stroke alerts are low‐technology interventions with a significant potential to improve acute stroke outcome. Because every second stroke alert in Sweden actually leads to reperfusion treatment Figure , it is nPublic stroke warning campaigns have been reported to have varying degrees of success (Mellon, Doyle, Rohde, Williams, & Hickey, The authors have no conflicts of interest. Click here for additional data file.


Spices and their bioactive components are more promising attractions for their inclusion in diet-based regimes to improve human health. These are sources of natural antioxidants and play an important role in the chemoprevention of diseases and aging. The aim of the current study was to explore the antioxidant potential of cinnamon; a widely used spice throughout the world.i.e., 0.05, 0.10, 0.15, 0.20 and 0.25%, to compare the antioxidant potential of the extract whereas, To acted as control and TBHA @ 0.1% was used as synthetic antioxidant in the oil samples. The oil samples were analyzed for rancidity check during storage (after every seven days for a storage period of four weeks).The current research was aimed to investigate the antioxidant potential of cinnamon extract. For the purpose, cinnamon sticks were procured from local super market, while palm oil was obtained from local oil industry. The resultant extract was analyzed for its antioxidant activity through total phenolic content (TPC), free radical scavenging activity (DPPH assay), and total antioxidant activity was measured by ferric reducing antioxidant power (FRAP) test. The shelf life of palm oil was checked by adding cinnamon extract in oil at different levels , To, TBHA, T1, T2, T3, T4, T5 were recorded as peroxide value , free fatty acids , TBA value  and iodine value . Overall, the statistical analysis indicated that all parameters regarding oil stability i.e., peroxide value (PV), free fatty acid content, (FFA) thiobarbituric acid (TBA) value and iodine value (IV) were significant with respect to treatments and storage.The results indicated that total phenolic contents (TPC); 1,1-diphenyl-2-picrylhydrazyl (DPPH) and ferric reducing antioxidant power (FRAP) values of cinnamon extract were as 355.01 ± 8.34 gallic acid equivalent per gram (mg GAE/g), 90.18 ± 2.12 (%) and 132.82 ± 3.12 (μmol/g), respectively. The oxidative parameters for treatments i.e.From the present study, it can be concluded that the cinnamon extract proved effective in reducing the lipid oxidation of palm oil and it can be successfully used in place of synthetic antioxidants in food preparations. The bioactive compounds of essential oils found from CZ bark are trans-cinnamaldehyde, eugenol and linalool, which are representative of almost 82.5% of total compounds [Novel health care strategies of the millennium have illuminated functional foods as one of the promising therapeutic tool to curb various lifestyle related maladies. In the recent era health enhancing properties of specific foods have been documented with antioxidant and anti-inflammatory properties . Spices cinnamon . The bioompounds . It possompounds –9. Moreoompounds –12.Oxidation reactions generates free radicals that start off chain reactions. This spontaneous process of oxidation causes rancidity and food spoilage. Moreover, oxidative stress leads to many lethal diseases in humans like cancer . CurrentPalm oil is an important part of human and animal diet and plays key role in food processing industry. Its use in the commercial food industry is widespread owing to low cost and frying stability. One of the important quality parameters in oil refining industry is low level of free fatty acid (FFA) content and oxidative products . HoweverCinnamon sticks were procured from Metro cash and carry, Faisalabad. They were dried in an oven at 55 °C till the constant moisture content of 8.5%, crushed using electrical grinder, refined & bleached. Deodorized (RBD) palm oil was obtained from United Ghee Industries Ltd., Faisalabad. All chemicals were of analytical grade and were purchased from Sigma-Aldrich ; Merck  and Fluka . The powdered sample were subjected to solvent extraction using ethanol.Solvent extraction was done following the method used by Al-Juhaimi and Ghafoor  with modw/v). The final mixture volume was maintained up to 7 mL with deionized water. After incubation for 2 h in dark at room temperature, absorbance of the resulting blue color was measured at 765 nm using UV-Visible spectrophotometer. Gallic acid was used as a standard for the calibration curve. The phenolic content was determined as gallic acid equivalents using the linear equation based on the calibration curve.Total phenolic contents of cinnamon extract were determined spectrophotometrically following Folin-Ciocalteu method described previously,  with min(SA) was calculated as follows:The free radical scavenging activity of the extracts was determined by using 1, 1-diphenyl-2-picrylhydrazyl or DPPH . The scaTotal antioxidant activity was measured by ferric reducing antioxidant power (FRAP) assay of Benzie and Strain .i.e., 0.05, 0.10, 0.15, 0.20 and 0.25%, to compare the antioxidant potential of the extract, whereas, To acted as control and TBHA @ 0.1% was used as synthetic antioxidant in the oil samples.The shelf life of palm oil was checked by adding cinnamon extract in oil at different levels To check the antioxidant potential of the extract on the oil stability, following chemical analysis were performed. Peroxide value was measured through procedure No. Cd 8–53 . Free faAll determinations were carried out in triplicate and data was reported as mean ± standard deviation. Data obtained for the different parameters was analyzed using the analysis of variance (ANOVA) technique and the least significance difference (LSD) test was used to compare the means, according to the procedure described by Steel et al. .The antioxidant activity of cinnamon extract was investigated using total phenolic content (TPC), free radical scavenging activity (DPPH assay), and ferric reducing antioxidant power (FRAP) test and the values were recorded as 355.01 ± 8.34 mg GAE/g, 90.18 ± 2.12 (%) and 132.82 ± 3.12 (μmol/g), respectively., To, TBHA, T1, T2, T3, T4, T5 was recorded as 2.61 ± 0.07, 2.42 ± 0.08, 2.57 ± 0.05, 2.56 ± 0.03, 2.54 ± 0.02, 2.54 ± 0.01, 2.46 ± 0.06 (Table o, TBHA, T1, T2, T3, T4, T5 were observed as 0.601 ± 0.05, 0.522 ± 0.02, 0.580 ± 0.07, 0.572 ± 0.03, 0.56 ± 00.07, 0.552 ± 0.03, 0.536 ± 0.05. The free fatty acid content decreased as the concentration of extract was increased. The effect was more pronounced for the synthetic antioxidant BHA, yet cinnamon extract at 0.25% had almost similar effect (Table In the current study the mean squares relating to PV, FFA, TBA and IV of palm oil associated with treatments and storage have been discussed in Table o, TBHA, T1, T2, T3, T4, T5, respectively (Table o, TBHA, T1, T2, T3, T4 and T5. Concerning the effect of storage, the data exhibited differences among storage days showing a little decline from 0 to 28th day. The values were 53.80 ± 2.12, 53.73 ± 2.96, 52.62 ± 2.63, 52.21 ± 2.12 and 51.90 ± 2.38 for 0, 7, 14, 21 and 28 days, correspondingly.The palm oil was also analysed periodically for TBA value during 28 days storage. The mean values obtained with respect to TBA value in fresh oil samples was 18.6 ± 0.73, whilst at 7, 14, 21 and 28 days the value increased gradually to 19.4 ± 1.45, 20.6 ± 1.56, 21.8 ± 1.56 and 24.7 ± 0.98. The means pertaining to treatments were observed as 22.4 ± 1.45, 20.1 ± 0.73, 21.8 ± 0.42, 21.2 ± 1.56, 20.7 ± 0.48, 20.5 ± 0.59, 20.2 ± 0.91, for TFicus exasperata leaves to improve the quality and stability of palm oil. They concluded that use of Ficus leaves enhanced the storage stability of palm oil. Liang et al. [p-anisidine value, free fatty acids, thiobarbituric acid-reactive substances and concluded that the natural antioxidants are better in reducing the oxidation damage to the oil as compared to synthetic one. The results of current research are in accordance with the findings of Zia-ur-Rehman et al. [The reactive oxygen species may cause deterioration of lipid base foods during processing and storage. They produce off flavors and odors consequently reducing the shelf life and marketability of foods. In the past, synthetic antioxidants were extensively used to prevent lipid oxidation, however recently their use in food is governed by strict regulations owing to their detrimental effects on human health. Today consumers are more conscious about their health and demand food that is more like fresh, resultantly this leads to search for antioxidants from natural sources. In this connection, spices are lavish source of polyphenolic compounds that possess strong antioxidant capacity and can be potentially used to substitute the synthetic antioxidants from food products and confer some additional health effects. The results of the present study regarding TPC, DPPH and FRAP assay were in accordance with the finding of Thakur et al. . They prg et al.  determing et al.  exploredn et al. . They exn et al.  concludeBoth natural and synthetic antioxidants may inhibit or postpone the process of fat oxidation. An antioxidant can play important role in the prevention of oxidation as by acting as reducing agent, free radical scavenger, chelator and as singlet oxygen scavenger . A numbeEssential oils and eugenol from cinnamon bark has exhibited very strong in vitro antioxidant activities, reducing nitrotyrosine generation thus decreasing the peroxy nitrite prompted lipid oxidation . The volDespite the proven antioxidant potential of the cinnamon extract further research is suggested to underpin recent analytical and biological evaluation approaches in order to gain a deeper understanding of possible antioxidant and health effects. Evaluation of cinnamon extract as antioxidant under different processing and storage conditions appears to be an appreciated future objective.The edible oils can be degraded through oxidation and decomposition of oxidation products, resulting in generation of alcohols, ketone, aldehyde, acids and hydrocarbons. They can badly affect the appearance, quality and palatability of a food product by creating obnoxious off-flavors and off-odors in food and hereafter affect safety, nutritional value, healthfulness, flavor, color and texture of different foods. These undesirable episodes of oxidation process can be inhibited by various ways including reduction of excess oxygen, and addition of oxidation inhibitors or antioxidants. There is growing trend for the use of natural antioxidants, for food ingredients preservation in recent years due to toxicity problems associated with synthetic antioxidants. The main purpose of the present research was to probe the antioxidant capacity of cinnamon spice. Moreover, the reducing ability of cinnamon extract on lipid oxidation was investigated after addition of extract in palm oil at different concentration levels along with synthetic antioxidant and control for comparison purposes. From the present study, it can be concluded that cinnamon extract has strong antioxidant potential and it can be successfully used in place of synthetic antioxidants in food preparations to improve human health and nutrition by reducing the production of hazardous oxidative derivates and free fatty acids.


However, the timing and mechanisms of changes in oxygen utilisation, associated with training beyond one year are not known. This study tested the hypothesis that in women aged 40–60 years performing the same current training load; systemic O2, and VL, GAST, and PFC HHb were measured simultaneously.Thirteen STT and 13 LTT participants performed RI and SWCL conditions on separate days. VO2peak was higher in LTT compared to STT, and VO2 was higher in LTT at each relative intensities of 25%, 80% and 90% of VTP in SWCL. HHb in the VL was significantly higher in LTT compared to STT at peak exercise (4.54 ± 3.82 vs 1.55 ± 2.33 μM), and at 25% (0.99 ± 1.43 vs 0.04 ± 0.96 μM), 80% (3.19 ± 2.93 vs 1.14 ± 1.82 μM) and 90% (4.62 ± 3.12 vs 2.07 ± 2.49 μM) of VTP in SWCL.VO2, and HHb in the VL at peak exercise, and sub—VTP exercise. These results indicate that in women 40–60 years old, systemic and muscle O2 utilisation continues to improve with ET beyond two years.The additional (12.9 ± 9.3) years of ET in LTT, resulted in higher VO The results indicated that trained, compared to untrained women had a better matching of O2 delivery to utilisation during moderate intensity exercise with a higher HHb amplitude reported in the trained compared to the untrained women at 90% VTP.Continuous wave near-infrared spectroscopy (NIRS) systems provide a more direct method of investigating changes in muscle and prefrontal cortex (PFC) deoxyhaemoglobin (HHb) during exercise , 19. Theng women –22 and Eng women  as well ng women , 22. Howng women . That st2Hb, tHb, TSI) occur during heavy exercise (in young men) ) was applied. This formula derived from data from Duncan et al. , with VO2 higher in LTT. There was a significant main effect with a large effect for intensity , with VO2 higher in LTT. There was no significant group x intensity interaction  (For relative VO= 0.415] .2 (L. min-1) in the RI, there was a significant group main effect with a medium effect size , with VO2 higher in LTT. There was a significant main effect with a large effect for intensity , with VO2 higher in LTT. There was no significant group x intensity interaction  (For absolute VO= 0.078] .2 (mL. kg-1. min-1) in the SWCL, there was a significant group main effect with a large effect size , with VO2 higher the LTT. There was a significant main effect with a large effect size for intensity , with VO2 higher in LTT. There was a significant group x intensity interaction , with VO2 being higher in LTT compared to the STT at 25%, 80% and 90% VTP (For relative VO 90% VTP .2 (L. min-1) in the SWCL, there was a significant group main effect with a large effect size , with VO2 higher the LTT. There was a significant main effect with a large effect size for intensity , with VO2 higher in LTT. There was a significant group x intensity interaction , with VO2 being higher in LTT compared to the STT at 25%, 80% and 90% VTP by 0.1 L. min-1, 0.2 L. min-1 and 0.4 L. min-1 respectively, and the difference between the groups widening as the intensity increased  = 8.630; p = 0.007; ɳƤ2 = 0.273; β = 0.803], with VE being higher in LTT. There was a significant main effect for intensity with a large effect size , with VE being higher in LTT. There was no significant group x intensity interaction  , with VE being higher in LTT. There was a significant main effect for intensity with a large effect size , with VE being higher in LTT. There was a significant group x intensity interaction , with VE being higher in LTT compared to the STT at 25% 80% and 90% VTP, with the difference between groups increasing as intensity increased  = 11.958; p = 0.002; ɳƤ2 = 0.342; β = 0.912], with VT being higher in LTT. There was no significant main effect for intensity . There was a significant group x intensity interaction , with VT being higher in LTT compared to the STT at 90% VTP, with the difference between groups decreased from 90% TP to peak exercise  = 6.471; p = 0.018; ɳƤ2 = 0.220; β = 0.683], with VT being higher in LTT. There was a significant main effect for intensity with a large effect size , with VT being higher in LTT. There was a significant group x intensity interaction , with VE being higher in LTT compared to the STT at 25% 80% and 90% VTP, with the difference between groups widening as intensity increased  = 17.144; p < 0.001; ɳƤ2 = 0.427; β = 0.977], with BF being higher in LTT. There was no significant group x intensity interaction   = 0.851; p = 0.366; ɳ= 0.505] .Ƥ2 = 0.049; β = 0.180]. There was a significant main effect for intensity with a large effect size , with BF being higher in LTT. There was no significant group x intensity interaction   = 1.173; p = 0.290; ɳ= 0.056] .Ƥ2 = 0.160; β = 0.479]. There was a significant main effect for intensity with a large effect size , with HR being higher in LTT. There was no significant group x intensity interaction   = 3.993; p = 0.059; ɳ= 0.137] .Ƥ2 = 0.009; β = 0.063]. There was a significant main effect for intensity with a large effect size , with HR being higher in STT. There was no significant group x intensity interaction   = 0.128; p = 0.726; ɳ= 0.229] .Ƥ2 = 0.111; β = 0.370]. There was a significant main effect for intensity with a large effect size , with RPE being higher in LTT. There was no group x intensity interaction   = 2.882; p = 0.103; ɳ= 0.5.8] .The HHb changes in the VL, GAST and PFC in RI followed the expected pattern . The HHbƤ2 = 0.238; β = 0.728], with HHb being higher in LTT. There was a significant main effect for intensity with a large effect size , with HHb being higher in LTT. There was no significant group x intensity interaction   = 7.176; p = 0.013; ɳ= 0.059] .Ƥ2 = 0.186; β = 0.592], with HHb being higher in LTT. There was a significant main effect for intensity with a large effect size , with HHb being higher in LTT. There was a significant group x intensity interaction , with HHb being higher in LTT compared to the STT at 25%, 80% and 90% VTP, with the difference between groups increasing as intensity increased  = 5.243; p = 0.032; ɳncreased .Ƥ2 = 0.002; β = 0.056]. There was no significant main effect for intensity . There was no group x intensity interaction   = 0.052; p = 0.821; ɳ= 0.064] .Ƥ2 = 0.000; β = 0.051]. There was no significant main effect for intensity . There was no group x intensity interaction   = 0.005; p = 0.944; ɳ= 0.437] .Ƥ2 = 0.001; β = 0.054]. There was a significant main effect for intensity with a large effect size , with HHb being higher in LTT. There was no group x intensity interaction   = 0.033; p = 0.857; ɳ= 0.059] .Ƥ2 = 0.000; β = 0.050]. There was no significant main effect for intensity . There was no group x intensity interaction   = 0.000; p = 0.999; ɳ= 0.085] .2, and multiple local muscle and PFC HHb during exercise in older women closely matched for current training load. It was hypothesised that VO2 and HHb would be higher in LTT compared to the STT at peak exercise, but similar when exercising at 25%, 80% and 90% VTP. The VO2 was significantly higher in LTT compared to the STT at peak exercise and at 25%, 80% and 90% VTP. Deoxyhaemoglobin (HHb) in the LVL was also significantly higher in LTT compared to the STT at peak exercise and at 25%, 80% and 90% VTP in the LV only. No difference in HHb was found for GAST or PFC at any intensity. A primary factor influencing VO2 and HHb during exercise is current training status, with greater VO2 and HHb in higher trained compared to less trained counterparts [This current study is unique in that it describes VOterparts , 2, 21. 2peak than STT older women. The absolute VO2peak values of 2.6 ± 0.4 L · min-1 for the LTT and 2.1 ± 0.4 L · min-1 for the STT are higher than reference values for untrained women of this age [2peak could be the primary physiological mechanism reflective of performance in older adults. However, they are in contrast to a dose response relationship existing between ET and improved VO2peak [2peak are achieved within 12 months of ET [2peak to increase beyond two years of regular ET. A possible reason for the conflict between the current VO2peak measures and others is that other studies have not investigated the changes in VO2peak beyond 12 months in older adults [This current study supported the hypothesis that, when matched for current training load, LTT older women have a significantly higher VOthis age , therefothis age , 51, 52 hs of ET . Our resr adults , rather r adults , 9.2peak. This suggests that a higher VTP as a percentage of VO2peak did not influence the higher VO2peak in LTT compared to the STT. This supports Marcell et al. [2peak is not a determining factor in aerobic exercise performance.As expected, absolute VTP values were significantly higher in the LLT compared to the STT, however, this difference was not seen when expressed as a percentage of VOl et al.  who repo2 (L · min-1) was significantly higher in LTT compared to the STT while cycling at a Power calculated as 25%, 80% and 90% of that at VTP. Moreover, as the intensity increased the difference between the groups changed from 0.1 L · min-1 at 25%, to 0.2 L · min-1 80% and 0.04 L · min-1 at 90% (as indicated by the ANCOVA interactions). These results support Dogra et al. [2 values in trained compared to untrained older women during constant load cycling at 90% VTP. This suggests that while exercising at the same relative intensity, those who are longer trained have improved O2 utilisation during sub-VTP constant load exercise.During the SWCL, contrary to our hypothesis, VOa et al.  who repoE in LTT at peak exercise and during the SWCL was produced by increased VT, not BF. These findings are in contrast to the response in typically healthy adults performing strenuous exercise, where VT plateaus once it reaches ~ 50–60% of vital capacity and further increases in VE result exclusively form increases in BF [Emax following ET have been reported in older women [E and VT at peak and sub—VTP intensities. These results indicate that the higher VE of the LTT was produced from breathing larger volumes of air (VT) rather than increases in BF. These higher volumes could be result of the LTT having more compliant connective tissue of the lungs and or better respiratory muscle function as a result of their additional years of training [An unexpected finding of this current study was the higher Ves in BF . While ier women , 55–57, training .2 delivery during SWCL exercise. Others have reported faster HR dynamics in trained compared to untrained older women, [As expected, peak HR was not different between groups during the RI. This supports other research that indicates that peak HR is a function of age, not training , 57. A dr women, , 21, howr women, . In the The rating of perceived exertion provides an additional valid method of evaluating effort based on subjective sensation . The SWC2 in the VL. A further unique component of the current study was the simultaneous measurement of HHb in the RVL and LVL. Results indicated that both legs utilised the same amount of O2 during exercise, and that the devices were measuring consistently.A unique component of this study was the simultaneous measurement of deoxyhaemoglobin (HHb) in multiple skeletal muscle and PFC. The results supported the hypothesis in that significantly higher absolute HHb were observed in the VL in the LLT compared to the STT at 90% VTP and peak exercise. However, the significantly higher HHb would be observed in the VL in LTT at each relative SWCL intensity  is contrary to the hypothesis. This suggests that regardless of exercise intensity, compared to STT, LTT utilise higher amounts of O2 delivery and utilisation (relationship between the time constants of HHb and VO2) in the VL during moderate exercise. While a peak exercise test was conducted in order to calculate the sub-maximal intensity, the authors did not report HHb values during the peak test. However, the higher absolute changes in HHb in the trained compared to untrained group during sub—VTP constant load exercise support the current study.Previously, only three studies have investigated HHb in exercising muscles of older women. Two compared young with old; one was unable to provide data due to technical difficulties , and theObserving no difference in HHb between the groups at GAST during the RI and SWCL are contrary the hypothesis. Oxygenation in the GAST during exercise has only ever been reported on men during treadmill running . The inv2 extraction, and lower percentages of Type IIa and IIb fibres which have a low perfusion pressure and rate O2 extraction [Differences in fibre structure between the GAST and VL may help explain the differences in HHb response between the muscles. That is, compared to the VL, the GAST has a higher percentages of Type 1 fibres which during exercise have a high perfusion pressure and rate of Otraction , 62. Furtraction . Age-reltraction . If thisOur observation of no difference in HHb between the STT and LTT groups in the PFC during the RI and SWCL support the hypothesis that the difference in training years between the groups in the current study would not influence PFC during exercise. The current study is unique in measuring the effect of ET on PFC HHb in healthy women, and supports similar findings in men . Further2peak values of the LTT compared to the STT indicate that improvements in the integration of the pulmonary, cardiovascular and muscular systems to uptake, transport and utilise O2, do not plateau after 12 months of aerobic ET as has been suggested [2 to maintain metabolic processes begins with increased pulmonary ventilation. A full description of the processes involved in ventilation and respiratory mechanics is beyond the scope of this current study, however, up to a point increases in VE result from increases in either VT or BF, or both [E result solely from increases in BF [E in LTT resulted solely from increase in VT not BF. The effects of age and ET on respiratory control of older women is relatively unknown, with interventional studies required before suggesting a relationship exists. However, the current study supports the proposal that higher levels of physical fitness in older women might alter typical age-related increases in ΔVE / ΔVCO2 [The higher VOuggested . During  or both , where fes in BF . In cont / ΔVCO2 .2 extraction. As stroke volume and cardiac output were not measured in the current study, it is not possible to determine any influence on the higher VO2peak of the LTT. However, as HR was not different between groups at any intensity during both conditions, and as maximum HR is not trainable, this indicates both groups were closely matched, and were exercising at the same relative intensity during the SWCL.Cardiac output (stroke volume x HR) provides the pressure and flow for oxygenated blood to reach exercising muscles for peripheral O2 utilisation as a wider a-vO2diff in older women following ET [2, below VT, lactate and metabolic needs plateau [2 uptake at this intensity is unlikely to influence peripheral HHb. Further, hyperoxia during exercise increases VO2 and arterial O2 saturation, while muscle HHb is not effected [2, the current study would have potentially observed significantly higher HHb in the GAST.The higher HHb in the VL in LTT in this current study support others who have reported improved peripheral muscle Oowing ET –6. This owing ET –69. Whil plateau , thus, aeffected , even ateffected . MoreoveMonitoring of blood flow and muscle activation between the VL and GAST require further investigation, and would assist in determining the reasons for the differences in deoxyhaemoglobin patterns between muscles.One potential limitation of this current study was the difference in the average age of the two groups, with the average age of the STT being four years older than the LTT, however, there was no group difference in peak HR, which is, predominately determined by age, providing support for the groups being well matched. A second limitation could be related to the average training load of LTT being higher than the STT. This was controlled for as best as possible through the recruiting process, and when tested was not significantly different. Nonetheless average weekly training loads were factored into the analysis as a covariant. A third limitation was that adipose tissue thickness was higher in STT compared to LTT. However, while high levels of adipose tissue may affect NIRS measures, all participants were within the recommended ranges (< 34 mm).2 utilisation during and following prolonged ET.This current study has implications for endurance trained older women, in that beyond the typical systemic and peripheral adaptations that accompany initial regular ET, prolonged ET provides further positive systemic and intramuscular adaptations. Future research should include training studies of more than 12 months duration and include measures of the systemic, central, and peripheral components of O2 at peak and constant load sub—VTP exercise. Further, these adaptations are concomitant with improvements in HHb in the VL during peak exercise and SWCL exercise below VTP. However, this higher deoxyhaemoglobin pattern is not observed in the GAST or PFC at any exercise intensity.It is concluded that that in women aged 40–60 years, continued regular ET beyond two years can significantly improve VO


The antioxidant potential of the essential oil from M. alternifolia was evaluated by the DPPH  method, thiobarbituric acid reactive species (TBARS) assay, and the hydroxyl radical scavenging activity method. The essential oil from M. alternifolia was able to reduce DPPH with an EC50  of 48.35 μg/ml, inhibit the lipid peroxidation with an IC50 (50% inhibitory concentration) of 135.9 μg/ml, and eliminate hydroxyl radicals with an EC50 of 43.71 μg/ml. Antimicrobial screening, minimum inhibitory concentration, and minimum bactericidal concentration assays showed that the essential oil from M. alternifolia inhibited strongly the growth of different types of microorganisms, including Escherichia coli, Staphylococcus aureus, Pseudomonas aeruginosa, Penicillium italicum Wehmer, and Penicillium digitatum Sacc. Thus, the essential oil of M. alternifolia possesses antioxidant and antimicrobial activity and could be suitable for use as a natural preservative ingredient in food, agriculture, and pharmaceutical industries.The Melaleuca alternifolia  belongs to the Myrtaceae family [ M. alternifolia has also been gradually introduced in southern China. The essential oil of M. alternifolia is derived by steam or hydrodistillation from M. alternifolia as secondary metabolites and is monoterpene-rich, volatile oil characterized by a strong odor [In the last decade, research into essential (volatile) oils has received increasing attention from both industrial and academic sectors because of the growing interest in green consumerism and the need for alternative techniques to assure the quality and safety of perishable foods . The plae family , which ie family . The plae family –7. In reong odor , 9. M. alternifolia, also known as tea tree, has been investigated as an alternative antimicrobial agent [ M. alternifolia. The purpose of the present study was to evaluate the in vitro antioxidant and antimicrobial activity of the essential oil from M. alternifolia.Several studies conducted on natural plant essential oils have indicated that these oils may be used as antimicrobial agents and have potential use in industrial applications –12. M. aal agent ; howeverMelaleuca alternifolia  samples were collected in four locations of Zhaoqing City of Guangdong Province at altitudes between 300 and 600 m. Samples were dried in well-ventilated spaces away from sunlight. Samples were then placed at 50°C to dry the samples to a constant, dry weight and disintegrated through a 90-mesh sieve. Air-dried plant materials were hydrodistilled using a Clevenger-type apparatus. The composition of M. alternifolia essential oil was analyzed in the laboratory using Hewlett-Packard 6890 gas chromatograph equipped with a cross-linked 5% PH ME siloxane Hewlett-Packard-5MS capillary column , coupled to a Hewlett-Packard 5972A mass spectrometer . The GC operating conditions were as follows: helium as carrier gas with a flow rate 2.0 ml/min; column temperature programming from 60°C to 275°C at 4°C/min; injector and FID detector temperatures of 215 and 275°C, respectively. The MS operating parameters were as follows: ionization potential, 70 ev; resolution, 1000; ion source temperature, 250°C. Identification of components was based on GC retention indices and the fragmentation patterns of the mass spectra with those of authentic samples, as well as the NIST 98 and HPCH 2205 GC–MS libraries. Relative percentage amounts were obtained directly from GC peak areas. The composition of the essential oil from M. alternifolia is presented in μM DPPH dissolved in methanol. After incubation for 30 min at room temperature, the absorbance was measured against a blank at 517 nm. The antioxidant activity was determined using the following equation:  = (Ablank − Asample/Ablank) × 100. The extract concentration providing 50% elimination (EC50) was calculated from the graph plotting inhibition percentage against extract concentration. Tests were carried out in triplicate.This spectrophotometric assay uses the stable 2,2-diphenylpicrylhydrazyl (DPPH) radical as a reagent . One milg for 5 min. Subsequently, 1.0 mL of the supernatant was mixed with 3.0 mL of a 0.67% TBA solution in 50% glacial acetic acid. The mixture was heated in a boiling water bath for ~30 min. The reaction mixture was centrifuged at 2000g for 5 min if the solution became turbid. Finally, the absorbance was measured at 535 nm. A decrease in absorbance indicates an increase in antioxidant activity. The antioxidant activity was determined using the following equation:  = (Ablank − Asample/Ablank) × 100. The extract concentration providing 50% inhibition (IC50) was calculated from the graph plotting inhibition percentage against extract concentration. Tests were carried out in triplicate.The thiobarbituric acid reactive substances (TBARS) method was used as described by . One mil2PO4-KOH buffer, pH 7.4), 0.1 mL of various concentrations of the samples, 0.2 mL EDTA (1.04 mM) and 200 μM FeCl3 (1 : 1 v/v), 0.1 mL of H2O2 (1.0 mM), and 0.1 mL ascorbic acid (1.0 mM) was incubated at 37°C for 1 h. One milliliter of thiobarbituric acid (1%) and 1.0 mL of trichloroacetic acid (2.8%) were added and the solution incubated at 100°C for 20 min. After cooling, the absorbance was measured at 532 nm. The antioxidant activity was determined using the following equation:  = (Ablank − Asample/Ablank) × 100. The extract concentration providing 50% elimination (EC50) was calculated from the graph plotting inhibition percentage against extract concentration. Tests were carried out in triplicate.The hydroxyl radical scavenging activity was measured by the method described in . The reaEscherichia coli ATCC25922 (E. coli), Staphylococcus aureus ATCC25923 (S. aureus), Pseudomonas aeruginosa ATCC27853 (P. aeruginosa), Penicillium italicum Wehmer , and Penicillium digitatum Sacc. (P. digitatum Sacc.) were obtained from the China Center for Type Culture Collection . M. alternifolia and placed on the surface of solid media plates previously inoculated with the different microorganisms (6.0 logCFU/mL) tested in this study. P. italicum Wehmer and P. digitatum Sacc. were cultured at 28°C and 120 rpm for 48 h, whereas E. coli, S. aureus, and P. aeruginosa were cultured at 37°C and 120 rpm for 24 h. The size of the halo for each microorganism was recorded by measuring the zones of growth inhibition surrounding the disks. Individual samples were examined in triplicate. The size of the halos is presented as means ± standard deviation.Paper disks with 6 mm diameter were soaked with 0.1 mL of the essential oil from M. alternifolia essential oil was added to glass tubes to yield final sample media concentrations at 0.2 to 48 mg/mL for the potato dextrose and Mueller Hinton broths, respectively. The control sets were run simultaneously without the addition of an antimicrobial agent. An appropriate volume of inoculum (6.0 logCFU/mL) was added to the media to give an approximate final cell concentration of 4.0 logCFU/mL. P. italicum Wehmer and P. digitatum Sacc. cultures in potato dextrose broth were incubated at 28°C and 120 rpm for 48 h, whereas E. coli and S. aureus cultures in Mueller Hinton broth were incubated at 37°C and 120 rpm for 24 h. The essential oil concentration that yielded no visible growth for a tested species was considered the MIC for that particular species. Then, 0.1 mL suspension obtained from the above transparent tubes was spread onto either potato dextrose agar (PDA) or Mueller Hinton agar (MHA). After 48 h culturing at 28°C (fungi) or 48 h culturing at 37°C (bacteria), the concentration of inoculated suspension containing the lowest level of antimicrobial agent that showed no colonies on the plate was determined as the MBC.A broth microdilution method was used to determine the MIC and MBC. The potato dextrose broth and Mueller Hinton broth were prepared at twice the final concentration. M. alternifolia can be obtained easily from the hydrodistillation of M. alternifolia leaves, and the chemical composition is dependent on the extraction method used and crop region the samples are taken from [ M. alternifolia essential oil used in this study are presented in γ-terpinene (25.30%), and α-terpinene (12.70%) were the major constituents followed by 1,8-cineole (6.83%), ρ-cymene (4.23%), terpinolene (4.03%), limonene (2.50%), α-terpineol (2.35%), aromadendrene (1.75%), and δ-cadinene (1.41%). As alluded to above, the composition of M. alternifolia essential oil is influenced by several factors, including local, climatic, seasonal, and experimental conditions. Thus, the biological activities of each essential oil preparation will likely vary [The essential oil fromken from . The cheely vary . M. alternifolia essential oil antioxidant activity. Vitamin C, vitamin E, quercetin, and α-lipoic acid are four known natural antioxidants and were used as positive controls. The DPPH assay has been used widely in determining the free radical scavenging properties of plant extracts [ M. alternifolia essential oil > α-lipoic acid. Reactive oxygen species, in particular hydroxyl radicals, are implicated in oxidative damage to fatty acids, DNA, proteins, and other cellular components [ M. alternifolia essential oil are similar to the DPPH assay results. Here, the antioxidant power decreased in the order quercetin > vitamin C > M. alternifolia essential oil > vitamin E > α-lipoic acid . These data are consistent with previous observations that Melaleuca alternifolia essential oil possesses antibacterial and antifungal activity [ M. alternifolia terpenes is associated with their strong hydrophobicity. Here, the hydrophobic terpenes interact strongly with the membrane lipids of the pathogenic microorganisms, which affect the permeability of the membrane [ Candida albicans [bacteria , whereasre fungi . Becauseactivity . Accordimembrane . Among tmembrane , 27. Thealbicans . in vitro antioxidant properties of the essential oil from M. alternifolia. Because of its strong antibacterial and excellent protective features exhibited in antioxidant activity tests, this essential oil and extracts from the herbal parts of M. alternifolia represent a potential natural source that can be used freely in food, agriculture, and pharmaceutical industries as a culinary herb.In conclusion, our study is the first report describing the