People usually spent about 90% of their time indoors, which are probably more polluted than outside the buildings. High levels of volatile organic compounds (VOCs) are known as causes of sick building syndrome. The present study was designed to determine the quantitative effects of some plants to improve the quality of the environmental air.D. deremensis and O. microdasys were chosen for the present study. There is no report of using O. microdasys for cleaning the air from pollutants. So, in this study, the effectiveness of O. microdasys in air removing from pollutants was studied and compared with D. dermensis.O. microdasys plant can remove 2 ppm concentration benzene, toluene, xylene and ethylbenzene from air in test chambers completely after 48, 55, 47 and 57 hours, respectively. The removal rates of benzene, toluene, xylene and ethylbenzene (BTEX) from air in the test chambers were 1.18, 0.54, 1.64 and 1.35 mg/ m2d1, respectively.3, it contains 16, 8, 22 and 22 mg/m3 benzene, toluene, xylene and ethylbenzene, respectively. Using ten O. microdasys pots with the same size used in this study, can remove benzene, toluene, xylene and ethylbenzene totally after 36, 40, 30 and 39 hours.If an office containing 2.5 ppm of each of BTEX and had an approximate volume of 30 mThe authors recommended studying the efficiency of the plants for removal of BTEX from air at higher range of concentrations such as 20-30 ppm. People generally spend about 90% of their time inside the buildings such as houses, offices and factories (indoor). The indoor air is probably more polluted than that of the outside buildings air (outdoor). Therefore, the possible effects of air pollution on human beings are an international issue -4. VolatStaff welfare and productivity were improved by putting plants indoor ,16. SeveIt has been showed that one of the VOCs removal agents are microorganisms of the soil. But, there was not significant effect on the VOCs removal for the majority of air contaminants ,19. AlsoGodish and Guindon reported the effects of spider plants on the removal of formaldehyde up to 50%. They showed that the removal effects of the plants were not primarily via the plants leaves. As they concluded that it must be rather due to other factors such as moisture, soil, microorganisms or all of them . SchmitzAglaonema modestum, Chamaedoreae legans and Philodendron for cleaning the environment from benzene, toluene, xylene and n-hexane, which are used as industrial solvents for furnishings. They showed that the contaminants concentrations were decreased gradually to below the detection limits of the gas chromatograph (<20 ppb). They found that the pot size in cleaning of the air is in less importance [Burchett et al. examined the capacity of three plants named portance .Dracaena deremensis and Opuntia microdasy to improve the quality of the environmental air.Reduction of VOCs levels especially in indoor environments is vital to improve human health. So, the present study was designed to determine the quantitative effects of Benzene, toluene, ethylbenzene, xylene, methanol, acetone and acetonitrile (HPLC grade) were purchased from Merck. All SPME supplies were obtained from Supelco. A CAR/PDMS fiber (75 μm) was used for sampling benzene from air. For sampling, EconoGrab 10 L Tedlar Bag with polyproplyene fitting and gas sampling pump  were used. The Tedlar bags after each sampling were purged three times with nitrogen for reuse.D. deremensis and O. microdasys were chosen for the present study. The capability of D. dermensis for removing of air pollutants such as benzene and toluene was confirmed by Orwell et al (26). There is no report of using O. microdasys for cleaning the air from pollutants. So, in this study, the effectiveness of O. microdasys in comparison with D. dermensis in air removing from pollutants was studied. Three year old plants were transferred to 10 cm diameter pots. Then, the pots left for two months in the laboratory to get habit with the new conditions. The temperature was kept at 20°C ± 3 and the light period was 12/12 hours dark/light. The plants were watered each three days and the remaining water was allowed to drain from the pots. All plants were watered one hour before putting under exposure to gas mixture. The characteristic of the plants are listed in the Table The plants were placed in the sealed chambers. After an hour, gas treatment was imposed on the plants. The plants with about the same leaf areas were placed in the chambers. Three replicates of each plant were examined for the gas treatment. The empty chambers without plants were tested as control chambers. It was used to determine the losses caused by leakage, chemical reaction or adsorption to the chamber surface. Leaves area of the plants were measured at the end of the test. At the end of each experiment, the plant was extracted from the soil and the pot was placed in the chamber to examine the gas absorption capacity of the soil alone.3 were purchased. Their diameter mouths were 35 cm that ease putting the pots inside the chamber. Removable glass lids were prepared and sealed with a solvent free adhesive tape. Rubber septa were installed on the centre of the lids for benzene, toluene, ethylbenzene and xylene injection and air sampling. A 2.4 W fan, controlled with remote, was installed inside the chambers to accelerate atmospheric equilibration. After benzene, toluene, ethylbenzene and xylene loading in the chamber, the fan was turned on for one hour, which is necessary for equilibration. As well, one hour before each sampling the fan was turned on to equilibrate benzene, toluene, ethylbenzene and xylene inside the chamber. Solid phase microextraction (SPME) fiber, CAR/PDMS fiber (75 μm), was used for their extraction from the chamber. The fiber was put inside the chamber in contact with the air and left it for equilibration. Then, the fiber was extracted from the chamber and inserted into the injection port for desorption.Glass chambers with the internal volume of 0.05 mGas chromatography (Younglin series YL 6100) with flame ionization detector was used to determine benzene concentration in the air. A capillary column DB-5MS (J & W Scientific) fused silica  was used. The operating conditions were: hydrogen flow 30 ml/min., air flow 300 ml/min, helium flow 0.8 ml/min, injector and detector temperatures were 260, 280°C, respectively. The GC oven was held at 40°C for 1 min, it was then ramped at 15°C/min to 90°C and held for 4 min, and finally ramped at 10°C/min to 170°C/min and held for 4 min. For calibrations, appropriate amounts of benzene, toluene, ethylbenzene and xylene and methanol  were put into a 10 L Tedlar bag and after equilibration they were extracted using a CAR/PDMS fiber (75 μm film thickness). Standard curves were consistently linear with R2 > 0.98. Quantitation was based on peak area ratio, which is relation between their concentrations to methanol concentration.The plants were watered and left for one hour to drain the extra water. Then, the plants were placed one in each chamber and the lids were sealed by adhesive. Benzene, toluene, ethylbenzene and xylene were added to chambers at final concentration of each 2 ppm. The samples were taken from chambers in duplicate and BTEX concentrations were analyzed using a gas chromatograph equipped with flame ionization detector (GC-FID Younglin series YL6100). Calibrations were performed at the BTEX concentrations of 0.2, 0.3, 0.4, 0.5, 1, 1.5 and 2 ppm. Samples were taken daily by a CAR/PDMS fiber (75 μm) and then immediately were analyzed using GC-FID. At the end of first experiment, the plants were removed and the pots with the soil alone were replaced in the chambers, which were sealed and they were exposed to a new dose of BTEX. For better plant removal, the pots were watered well, and then the plants extracted gently removing as little soil as possible. Finally, the experiment was performed with empty chamber to test the leakage.Means (n = 3) and standard errors (SE) were calculated. Daily BTEX values obtained were subjected to ANOVA analysis. Differences between variables were reported as statically significant where p value was less than 0.05.2 = 0.9936, 0.9962, 0.9986 and 0.9956, respectively). BTEX concentrations could be decreased by by chemical destruction, absorption to the chamber and destruction by soil bacteria. To avoid the effects of these factors, the results of the plants were compared with the soil results and the differences between them were significant. Both D. deremensis and O. microdasys significantly reduced concentration of benzene, toluene, ethylbenzene and xylene in the chambers  in Tedlar bag air are shown in Figure  in Tedla Figures , 5.D. dermensis and O. microdasys plants were expressed based on leaf area, cleaned cubic meter of the air and both of them. Kim et al reported the ability of a few plants to remove volatile substances from indoor area [Some of the plants have the capacity to remove air concentrations of VOCs, which knows as phytoremediation. Various plants have different capacities, which in this study the removal ability of the oor area .O. microdasys was able to remove 2 ppm of benzene from air in the test chambers completely after 48 hours. However, D. dermensis could remove benzene from air in the test chambers completely after 105 hours. The removal rates of benzene in the test chambers were 3.2 and 1.46 mg/ m3d1 for O. microdasys and D. dermensis, respectively. Also, the removal rates of benzene from air in the test chambers were 1.18 and 0.52 mg/ m2d1 based on leaf area of O. microdasys and D. dermensis, respectively. On the other word, O. microdasys and D. dermensis abilities were 23.2and 10.6 mg/ m3m2d1, respectively. However, David Moerlein investigated the ability of S. trifasciata, F. robusta, C. seifrizii and D. dermensis in laboratory to reduce total air borne inorganic and organic particles 10 microns or smaller in diameter. The plants did not significantly affect total particles in rooms [1d1 that removed by the P. Domesticum plant [in rooms . Corenjoum plant . Their rO. microdasys was able to remove 2 ppm of toluene from air in the test chambers completely after 55 hours. However, D. dermensis could remove toluene from air in the test chambers completely after 120 hours. The removal rates of toluene in test chambers were 1.47 and 0.67 mg/ m3d1 for O. microdasys and D. dermensis, respectively. Also, the removal rates of toluene from air in the test chambers were 0.54 and 0.24 mg/ m2d1 based on leaf area of O. microdasys and D. dermensis, respectively. On the other word, O. microdasys and D. dermensis abilities were 10.7 and 4.86 mg/ m3m2d1, respectively. Yang et al used 28 different plant species to determine their removal abilities. They showed that these plants could decrease toluene and benzene concentrations in a closed jar [Four different plant species were studied by Yoo et al to determine their removal abilities. Their removal abilities for benzene and toluene was good,which confirmed the results of present study . HoweverJanet Craig, E. aureum, D. marginata, S. amate, S. petite, S. sensation and H. forsteriana were studied by Orwell et al, and they were more than that of O. microdasys and D. dermensis plants [The removal rates of benzene by the D. s plants .3d1 and 2.12 mg/ m3d1 for O. microdasys and D. dermensis, respectively. Also, the removal rates of xylene from air in test chambers are 1.64 mg/ m2d1 and 0.76 mg/ m2d1 based on leaf area of O. microdasys and D. dermensis, respectively. On the other word, O. microdasys and D. dermensis abilities were 32.03 and 15.36 mg/ m3m2d1, respectively. Orwell et al reported the removal ability of S. Sweet Chico and D. Janet Craig plants for toluene and xylene. S. Sweet Chico and D. Janet Craig plants were prior to the plants used in the present study for removing toluene from air. However, the ability of O. microdasys for removing xylene from the air was prior to S. Sweet Chico and D. Janet Craig plants [O. microdasys plant can remove 2 ppm concentration xylene from air in test chambers completely after 47 hours. However, D. dermensis can remove xylene from air in test chambers completely after 98 hours. The removal rates of xylene in test chambers are 4.42 mg/ mg plants .3d1 and 2.1 mg/ m3d1 for O. microdasys and D. dermensis, respectively. Also, the removal rates of ethylbenzene from air in test chambers are 1.35 mg/ m2d1 and 0.76 mg/ m2d1 based on leaf area of O. microdasys and D. dermensis, respectively. On the other word, O. microdasys and D. dermensis abilities were 26.45 and 15.22 mg/ m3m2d1, respectively.O. microdasys plant can remove 2 ppm concentration ethylbenzene from air in test chambers completely after 57 hours. However, D. dermensis can remove ethylbenzene from air in test chambers completely after 100 hours. The removal rates of ethylbenzene in test chambers are 3.65 mg/ mIn authors knowledge, there was not found any study work on the removal of the ethylbenzene so far.3, it contains 16, 8, 22 and 22 mg/m3 benzene, toluene, xylene and ethylbenzene, respectively. Using ten of O. microdasys pots with the same size used in this study, can remove benzene, toluene, xylene and ethylbenzene totally after 36, 40, 30 and 39 hours, respectively.If an office containing 2.5 ppm of each of BTEX and had an approximate volume of 30 mThe authors recommended studying the efficiency of the plants for removal of BTEX from air at higher range of concentrations such as 20-30 ppm.In conclusion, there are a few studies reported the ability of different plant species in removing organic chemicals from the air. Various plants have different abilities for removal of air borne inorganic and organic particles. The best plant is one with higher removal rate, which could be recommended to be used indoor especially in high air polluted area.VOCs: Volatile organic compounds; BTEX: Benzene, toluene, xylene and ethylbenzene; SPME: Solid phase microextractionThe authors declare that they have no competing interests.All authors precipitated in conception and design, generation of data, analysis of data, interpretation of data, drafting of manuscript, revision of manuscript and approval of the final draft.


The compounds were confirmed by physical and analytical data and screened for in vitro antioxidant activity by DPPH method, reducing power assay and total antioxidant capacity method. The cytotoxic activity of the compounds was also studied in HEp-2 cell line. The docking studies of the synthesized compounds were performed towards the key nucleoside dsDNA by using AutoDock vina 4.0 programme. All the isoxazole-substituted compounds have significant activities.A convenient synthesis of novel isoxazole-substituted 9-anilinoacridine derivatives  Acridine derivatives are used in medicine and have enormous potential as pharmaceutical agents due to their biological activities such as antimicrobial , antioxi1H NMR and 13C NMR (Bruker A VIII 500 MHz Spectrometer) spectra have been recorded. Mass spectra of the final compounds were recorded on a JEOL GC mate Mass Spectrometer. The synthetic procedure was explained in the All the organic solvents used were of high purity unless otherwise stated. The reactions were monitored by TLC on silica gel thin layer plates. Elemental analysis , melting points (in glass capillary tubes on a Veego VMP-1 Apparatus), IR spectra (Shimadzu 8400 FT-IR spectrometer), In a 250 mL round-bottomed flask 4.06 g (0.03 mole) of 4-aminoacetophenone was refluxed with 5.4528 g (0.0256 mole) of 9-chloroacridine in 80 mL of 2-butanol for 3 h. After completion of reaction the reaction mixture was allowed to cool to room temperature then it was poured into 150 mL of ice water. A precipitate formed was filtered by suction, washed with water, dried, and recrystallized from ethanol.The chalcones were synthesized by using general Claisen-Schimdt condensation. In a 100 mL flat-bottomed flask 25 mL of the 10% sodium hydroxide and 25 mL of ethanol were taken with a magnetic stirring bar, it was placed on the magnetic stirrer, and the stirring dial was adjusted to get a nice, even stirring action. To this 0.0115 mole of corresponding aldehyde was added, then 2.9952 g (0.0096 mole) of 1-[4-(acridin-9-ylamino)phenyl]ethanone was added at the last. The solution was allowed to stir for 8 h at room temperature. After completion of the reaction 100 mL of water was added, formed precipitate was filtered and washed three times with 50 mL of water each time to remove sodium hydroxide, dried, and crystallized from ethanol. All the compounds are characterized and confirmed by IR, NMR, and mass spectroscopy. All the values are agreed with the synthesized compounds.ν, cm−1) for compound 4a: 3269 (N–H), 3057-3000 (Ar C–H), 1647 , 1604 & 1516 (Ar C=C), 1226 (C–N), 759 (Ar C–H); ms : m/z 400.16 (M+); 1H NMR (in ppm): 6.65–8.02 , 7.90 and 7.56 , 11.21 ; 13C NMR (in ppm): 183.4, 153.8, 149.8, 142.8, 140.5, 136.1, 135.4, 131.7, 131.3, 130.4, 129.3, 129.8, 128.9, 127.8, 126.7, 126.7, 121.9, 119.3, 115.7.The yield of compounds is between 60 and 85%; IR . After completion of the reaction excess solvent was concentrated in vacuo at reduced pressure, residual solution obtained was cooled to room temperature, poured into ice cold water, neutralized with sodium hydroxide, and washed well with water to remove unreacted hydroxyl amine hydrochloride and excess of sodium hydroxide. Dried crude product yield was noted and recrystallized from ethanol. All the compounds are characterized and confirmed by IR, NMR and mass spectroscopy. All the values are agreed with the synthesized compounds.In a 25 mL beaker, 2.5 g (0.02 mol) anhydrous sodium acetate was dissolved in minimum amount of hot glacial acetic acid (10 mL) and it was added to a solution of 0.81 g (0.01 mol) hydroxyl amine hydrochloride in absolute ethanol (10 mL). A solution of 0.01 mol of the corresponding chalcones 28H19N3O: C, 81.34; H, 4.63; N, 10.16; O, 3.87. Found: C, 81.21; H, 4.42; N, 10.1; IR : 3340 (N–H), 2993 (Ar st C–H), 1600 & 1473 (Ar C=C), 1157 (Ar C=N), 1226 (C–O), 742 (Ar C–H); MS: m/z 413.15 (M+); 1H NMR (in ppm): 6.85–8.94 , 9.95 , 11.10 ; 13C NMR (in ppm): 98.4 (C of isoxazole), 114.8–143.1 (27 aromatic carbons).This compound is obtained as a yellow powder; m.p.: 170–172°C; Yield: 56%; Anal. calc. for CThe computer-simulated automated docking studies were performed using the widely distributed molecular docking software, AutoDock Vina 4.0. The proteins were extracted from protein database at the NCBI, that is, dsDNA octamer duplex (1XRW). The designed 9-anilinoacridine analogues were taken for prediction of 3D structure by using Cambridge software. The energy was minimized for flexible docking using Argus Lab, for the docking of ligands to protein active sites and for estimating the binding affinities of docked compounds by an advanced molecular docking program AutoDock Vina, version 4.0. Predicting the binding affinity and rank-ordering ligands in database screens was implemented by modified and expanded version of the ChemScore18 scoring functions, for use. Using standard precision (SP) mode of PyMOL software, docking studies were performed on planned compounds for synthesis. The structures of the compound and enzymes are shown in 4a–j, 5a–j were screened for antioxidant activity and short-term in vitro antitumor activity against Daltons Lymphoma Ascites (DLAs) cells. Many of the synthesized final compounds 5a–j have significant activities.Acridine derivatives possess a diverse range of pharmacological activities –16. HencμL of DPPH solution, 10 μL of each of the synthesized or standard solution was added separately in wells of the microtitre plate. The final concentration of the synthesized compounds and standard solutions used were 1000, 500, 250, 125, 62.5, 31.25, and 15.625, 7.812 μg/mL. The plates were incubated at 37°C for 30 min and the absorbance of each solution was measured at 490 nm, using ELISA reader. The percentage inhibition was calculated. IC50, which is the concentration of the sample required to scavenge 50% of free radicals.The assay was carried out in a 96-well microtitre plate. To 200 10 mL of each of the synthesized or standard solution was added separately in 1.0 mL of deionized water that were mixed with phosphate buffer (2.5 mL) and potassium ferri cyanide (2.5 mL). The mixture was incubated at 50°C for 20 min. Aliquots of trichloroacetic acid (2.5 mL) were added to the mixture, which was then centrifuged at 3000 rpm for 10 min whenever necessary. The upper layer of solution (2.5 mL) was mixed with distilled water (2.5 mL) and a freshly prepared ferric chloride solution (0.5 mL). The absorbance was measured at 700 nm. Ascorbic acid at various concentrations was used as a reference standard. Phosphate buffer (pH 6.6) was used as blank solution. The absorbance of the compounds taken was expressed as mean ± standard deviation. Increased absorbance of the reaction mixture indicates increase in reducing power.The antioxidant power of the compounds has been assessed with the phosphomolybdenum reduction assay according to Armatu et al. The assay is based on the reduction of Mo(VI)-Mo(V) by the compounds and subsequent formation of a green phosphate—Mo(V) complex at acid pH. Various concentrations compounds were combined with 3 mL of reagent solution . The tubes containing the reaction solution were incubated at 95°C for 90 min. Then the absorbance of the solution was measured at 695 nm using spectrophotometer against blank after cooling to room temperature. DMSO (0.3 mL) in the place of compounds was used as the blank. For reference, the various concentrations of ascorbic acid have been used.6 cells/mL. The drug dilutions were made with phosphate buffer saline, and the drug dilutions were further adjusted to required concentrations. The drug dilutions were then added to the Daltons Lymphoma Ascites (DLAs) cells and incubated at 37°C for 3 hours. At the end of 3 hours, trypan blue dye exclusion test was performed, and percentage viability and percentage cytotoxicity were calculated.Cells were collected, counted, and adjusted to 1 × 10in vitro antioxidant activity, all the synthesized chalcone and isoxazole-substituted 9-anilino acridines were screened by DPPH assay, reducing power assay, and TAC almost all the isoxazole-substituted derivatives have shown antioxidant activity, among these compounds 5a, 5c, 5d, 5f, 5i, and 5j having more potent antioxidant activity when compared to standard ascorbic acid. The IC50 values, which is the concentration of the sample required to increase 50% of reducing capacity of the compounds, are shown in For 5a–j were subjected to short-term study for in vitro cytotoxic activity against Daltons Lymphoma Ascites (DLA) cells. All the isoxazole substituted derivatives were shown significant cytotoxicity and the compound 5i has more potent cytotoxic activity. The results were shown in The synthesized final compounds For the docking of ligands to protein active sites and for estimating the binding affinities of docked compounds, an advanced molecular docking program, AutoDock Vina version 4.0, was used in this study. The designed analogues are docked towards the dsDNA octamer duplex in order to ascertain their anticancer activity. The analogues show best fit Root Mean Square Difference (rmsd) value of 0.000. in vitro anticancer activity when compared to the control. The molecular docking studies show a good correlation between their biological activities screened and autodock binding free energy. In particular compound 5i shows significant anticancer activity. Compound 5i have potent cytotoxic activity and is likely to be useful as drugs after further refinement. These derivatives will encourage helping to design future anticancer agents with therapeutic potentials.A series of novel isoxazole-substituted 9-anilino acridine derivatives have been synthesized. The synthesized compounds have significant antioxidant activity when compared with standard ascorbic acid and also showed significant cytotoxic activity by


Epimedium extract and its constituent icariin on peripheral nerve repair were investigated in a crush injury rat model. Animals were divided into four groups: sham, control, Epimedium extract, and icariin groups. At postoperative weeks 1, 2, 4, and 8, nerve regeneration and functional recovery were evaluated by sciatic functional index (SFI), nerve electrophysiology, nerve pinch test, and muscle wet weight. Results showed that at 2 and 4 weeks after surgery rats in the Epimedium group displayed a better recovery of nerve function than that in the icariin and control groups, with better recovery in the icariin group than in the control group. The nerve pinch test showed that nerve regeneration was greater in the Epimedium group and the icariin group as compared to the control group. In addition, the muscle wet weight in the Epimedium group was significantly improved when compared with the icariin group, and the improvement in the icariin group was better than that in the control group at 8 weeks after operation. Our findings suggest that Epimedium extract effectively promotes peripheral nerve regeneration and improves the function of damaged nerves.Effects of  Treatment of peripheral nerve injury is a major challenge in clinical practice. With advances in molecular biology and development of microsurgical techniques and tissue engineering, peripheral nerve repair procedures have been greatly improved . In the Epimedium, and so forth, could enhance the peripheral nerve regeneration in rats [Epimedium may be a major component promoting the peripheral nerve regeneration in the formula. Epimedium has been used in China to treat erectile dysfunction, postmenopausal syndrome, and osteoporosis for thousands of years [Epimedium [Epimedium, Shindel et al. have demonstrated that icariin can promote bone formation and has neurotrophic effects in both in vivo and in vitro experiments [Epimedium koreanum could promote recovery of muscle function after spinal cord injury in rats [Epimedium extract and its main ingredient, icariin, on peripheral nerve regeneration were investigated.In our previous studies, results showed that systemic administration of a traditional formula, which mainly contains the Radix Hedysari,  in rats . Thus, wof years . Icariinpimedium , 14. Baseriments . Tohda a in rats . In the ad libitum access to water and food. All efforts were made to minimize animal suffering and to reduce the number of animals used. All of the surgical procedures, experimental manipulations, and perioperative care were performed in strict accordance with the Chinese Guidelines for the Care and Use of Laboratory Animals and were approved by the Hospital Medical Ethics of Peking Union Medical Collage.A total of 42 healthy adult male SD rats weighing 200–220 g (SPF grade) were purchased from the Beijing Unilever Animal Co., Ltd., and housed in the Animal Center of People's Hospital of Peking University. Animals were maintained in a specific pathogen-free (SPF) environment with controlled humidity and 12:12 h light-dark cycle and were given n = 6), control group (n = 12), Epimedium extract group (n = 12), and icariin group (n = 12). Rats were anesthetized with a single intraperitoneal injection of 2% pentobarbital solution (30 mg/kg). In the control, Epimedium extract, and icariin groups, the right sciatic nerve was exposed, clamped at 5 mm above the first branch of the nerve for 1 min, and marked with a 10-0 nylon microscopic suture in the epineurium under aseptic conditions. In the sham group, the right tibial nerve was only exposed, without nerve crush. Animals in the sham and the control group were intragastrically treated daily with 1 mL of distilled water. Rats in the Epimedium extract group and the icariin group were intragastrically fed daily with 1 mL of Epimedium extract and icariin, respectively.All the animals were randomly divided into 4 groups: sham group  were immersed in pure water at a volume ratio of 1 : 10 and boiled for 1 h. This was repeated once and the supernatant was collected by using a 200-mesh gauze filter. The collected supernatant was kept at room temperature overnight to remove sediments. It was then concentrated to 1000 mL by boiling and kept at 4°C for use. Five grams of icariin was dissolved in 1000 mL of pure water and kept at 4°C for use.The sciatic functional index (SFI) was measured at weeks 1, 2, 4, and 8 after surgery. A walking track box (50 cm in length) was made and a white paper was cut to the appropriate dimensions and placed on the bottom. The hindlimbs were dipped into carbon ink and each rat was permitted to walk down the box in order to record the bilateral footprints 4-5 times. The following parameters were determined: (1) footprint length (PL), defined as the distance from the heel to toe; (2) toe spread (TS), defined as the distance from the first to fifth toes; and (3) intermediary toe spread (IT), defined as the distance between the second and fourth toes. The right footprints (E) were recorded and measured, and the left footprints served as controls (N). Three parameters were calculated: print length factor (PLF) = (EPL-NPL)/NPL, toe spread factor (TSF) = (ETS-NTS)/NTS, and intermediary toe spread factor (ITF) = (EIT-NIT)/NIT. These parameters were utilized to calculate the Bain-Mackinnon-Hunter (BMH) sciatic function index (SFI) as follows: SFI = −38.3 (PLF) ± 109.5 (TSF) ± 13.3 (ITF) − 8.8. The recovery rate of SFI was calculated as a score of 0–100. Zero represents normal and 100 as no function.Epimedium extract, and icariin groups for nerve pinch test at postoperative week 1. Rats were anesthetized with a single intraperitoneal injection of 2% pentobarbital solution (20 mg/kg). The right sciatic nerve was exposed again, and a clamp was gradually applied from the distal tibial nerve to the proximal tibial nerve. Then, the pain reflex was observed, and the distance between the farthest reflective site and the crushed site was recorded. Experiments in this study were double blind.Six rats were randomly selected from the control, μm sections were obtained. The slices were dried at room temperature for 2 h, kept at 4°C, fixed in acetone at −20°C for 20 min, and washed thrice with 0.3% triton-PBS (5 min for each). These sections were blocked with 10% normal goat serum for 1 h and incubated overnight with mouse anti-rat GAP-43 monoclonal antibody  at 4°C. The slides were incubated with rabbit anti-mouse IgG-FITC antibody  at room temperature for 1 h after being washed thrice with 0.3% triton-PBS (5 min for each). Sections were observed under a fluorescence microscope.After nerve pinch test, animals were killed under anesthesia and sciatic nerves were collected from the 5 mm proximal crush point to the 20 mm distal point. Sciatic nerves were fixed in 4% paraformaldehyde for 6 h and then dehydrated in 15% sucrose for 8–12 h and 30% sucrose overnight. The nerves were embedded in OCT compound. At −20°C, 10 At week 8, rats were anesthetized and sciatic nerves were exposed. Electrophysiological recordings were performed in a quiet room with an ambient temperature at 22-23°C. For the detection of compound muscle action potential (CMAP), the recording electrodes were located on the central portion of the triceps and reference electrodes on the ipsilateral thigh muscle. Paraffin was applied around the neural stem to reduce fluid pathway conduction. CMAP was recorded after a stimulus with the intensity of 0.9 mA was given, using a pulse width of 0.1 ms and a frequency of 1 Hz. The latency of CMAP was recorded after stimulation of the nerve at crushed site and 30 mm proximal from the sciatic nerve, and the nerve conduction velocity  was obtained semiautomatically by dividing the distance between two stimulating sites by the difference in the onset latency.Experimental rats were sacrificed by overdose anesthesia after electrophysiological test. Bilateral triceps were carefully separated, collected, and weighted as the muscle wet weight. The ratio of right weight to left weight was calculated. A 5 mm muscle was harvested with a scalpel blade from the middle of the muscles, formalin-fixed, and paraffin-embedded for HE staining. The morphology of muscles was observed under a microscope.P less than 0.05 was considered statistically significant.Data were expressed as mean ± standard deviation. Comparisons of means among different groups were done using one-way analysis of variance (ANOVA). A value of Epimedium extract, and icariin groups was −7.54 ± 1.75, −86.95 ± 4.54, −85.98 ± 5.30, and −88.39 ± 7.64, respectively, at week 1; −5.88 ± 2.03, −70.08 ± 9.71, −56.65 ± 8.36, and −63.58 ± 9.23, respectively, at week 2; −6.32 ± 2.90 −27.50 ± 7.22, −18.61 ± 4.91, and −23.79 ± 3.03, respectively, at week 4; −6.15 ± 2.24, −12.71 ± 3.79, −10.20 ± 3.98, and −13.12 ± 2.94, respectively, at week 8. In the control, Epimedium extract, and icariin groups, SFI showed a gradual recovery of sciatic function, but the sciatic function was worse in the control group. At weeks 2 and 4, the neurological function was significantly improved in the Epimedium group and the icariin group as compared to the control group. At week 8, the SFI was comparable among these groups  and the icariin group (P = 0.026), but no significant difference was observed between the icariin group and the control group (P = 0.303)  .Epimedium extract and icariin groups at week 1 after operation . There was no significant difference between the Epimedium extract and the icariin groups (P = 0.17).The nerve conduction velocity of the sham, control, Epimedium extract and the icariin groups. The wet weight of left normal muscle in the sham, control, Epimedium extract, and icariin groups was 1.285 ± 0.098 g, 1.271 ± 0.088 g, 1.303 ± 0.138 g, and 1.307 ± 0.086 g, respectively, showing no significant difference between any two groups. The wet weight of right muscle was 1.283 ± 0.071 g, 0.853 ± 0.042 g, 1.075 ± 0.111 g, and 1.010 ± 0.109 g in the sham, control, Epimedium extract, and icariin groups, respectively. The recovery rate of wet muscle weight was 100.10 ± 5.10%, 67.26 ± 3.67%, 82.56 ± 4.35%, and 77.13 ± 3.79% in the sham, control, Epimedium extract, and icariin groups, respectively. The muscle wet weight and recovery rate of the control, Epimedium, and icariin groups were lower than those of the sham group. The wet muscle weight and recovery of the control group were lower than those in the Epimedium group and the icariin group, and the muscle recovery rate in the Epimedium extract group was better than that in the icariin group.The bilateral wet muscle weight of the sham group was comparable. Atrophy of the muscle was evident in the control group, however, as compared to the Epimedium extract, and icariin groups, muscle fiber staining was not uniform, and the muscle diameter was smaller than that in sham group. Muscle diameter of the control group was the smallest . Results demonstrated that Epimedium extract accelerated axonal growth, which is helpful to explain why the SFI in the Epimedium group was significantly improved at 2 and 4 weeks after operation as compared to the control group. Findings in nerve electrophysiological measurement, muscle wet weight measurement, and histological staining also supported that both Epimedium extract and icariin can exert protective effects on the motor function recovery and conductivity recovery in rats. It is presumed that icariin alone is effective to promote nerve regeneration, and may be one of the major components of Epimedium extract promoting the regeneration of peripheral nerves. However, the effects of Epimedium extract and icariin on the peripheral nerve regeneration are different, suggesting that components other than icariin in Epimedium extract may also facilitate the regeneration of peripheral nerves or coordinate this growth.The SFI most commonly employed is the BMH SFI formula –19. In tEpimedium extract on the peripheral nerve regeneration. This model is easy to establish and can be appropriately standardized, and it successfully reflects the restoration of nerve function. The sciatic nerve crush may produce a Sunderland type II injury, in which the myelin sheath and the axons are disrupted, but the basal lamina Schwann cell tubes remain intact [There are three animal nerve injury models frequently used in studies on the physiology and function of peripheral nerves: crush, transection, and graft/conduit , 21. In n intact . Thus, tEpimedium extract used in this study was determined according to our previous studies [Epimedium extract was determined by high-performance liquid chromatography. The average proportion of icariin is about 4.873 mg/mL. Thus, the concentration of icariin prepared in this study was identical in both Epimedium extract and icariin solution.The dose of  studies . The proEpimedium on nerve regeneration and growth, and the detailed mechanisms are still unclear. Further studies are warranted.Clinical and experimental studies have demonstrated that many TCM can promote peripheral nerve regeneration. For example, Wei et al. found that Hedysari extract (a TCM) prompted the regeneration of peripheral nerves in animal experiments . Hsiang


Thermoascus aurantiacus on solid-state fermentation was carried out in culture media containing sugar cane bagasse and orange bagasse in proportions of 30% and 70% (w/w) at 45°C for 4 days. PG obtained was purified by gel filtration and ion-exchange chromatography. The highest activity was found between pH 4.5 and 5.5, and the enzyme preserved more than 80% of its activity at pH values between 5.0 and 6.5. At pH values between 3.0 and 4.5, PG retained about 73% of the original activity, whereas at pH 10.0 it remained around 44%. The optimum temperature was 60–65°C. The enzyme was completely stable when incubated for 1 hour at 50°C. At 55°C and 60°C, the activity decreased 55% and 90%, respectively. The apparent molecular weight was 29.3 kDa, Km of 1.58 mg/mL and Vmax of 1553.1 μmol/min/mg. The presence of Zn+2, Mn+2, and Hg+2 inhibited 59%, 77%, and 100% of enzyme activity, respectively. The hydrolysis product suggests that polygalacturonase was shown to be an endo/exoenzyme.Polygalacturonases are enzymes involved in the degradation of pectic substances, being extensively used in food industries, textile processing, degumming of plant rough fibres, and treatment of pectic wastewaters. Polygalacturonase (PG) production by thermophilic fungus  Pectinases are a heterogeneous group of enzymes that hydrolyze the pectic substances present in plant material. The classification of pectinases is based on their mode of attack on the galacturan backbone of the pectin molecule, on specificity by substrate, or according to region of molecule where it acts .α-1-4 linkages in pectic acid and are of two types: endo-polygalacturonases , which act by hydrolysis of internal glycosidic bonds α-1-4 of polygalacturonic acid at random form, resulting in molecule depolymerization with release of oligogalacturonic acids, and exo-polygalacturonases  which hydrolyse alternate α-1-4 glycosidic linkages of polygalacturonic acid from the nonreducing end, releasing unsaturated mono- or digalacturonic acids [The polygalacturonases catalyze the hydrolysis of glycosidic This group of enzymes has been widely used in the food industry process such as clarification and viscosity reduction of fruit juices, preliminary treatment of grape juice for wine industries, tomato pulp extraction, oil extraction, and tea fermentation and in the textile industry in fibers degumming , 5.Aspergillus oryzae cultivation in solid-state fermentation (SSF) were more resistant to pH and temperature changes compared to those obtained by submerged fermentation (SmF). Moreover, Martin et al. [Thermomucor indicae-seudaticae showed higher thermostability in SmF than that in SSF. Thus, the present study aimed to purify polygalacturonase produced by thermophilic fungus Thermoascus aurantiacus in solid-state fermentation and compare its properties with those of the purified polygalacturonase produced by the same fungus in SmF, in a work reported by Martins et al. [In the literature, it has been reported that the type of fermentation influences the enzymes properties, such as thermostability and tolerance to pH variations , 7. Accon et al.  reporteds et al. .Thermoascus aurantiacus CBMAI756 was used. The strain is deposited in the Coleção Brasileira de Microrganismos de Indústria e Meio Ambiente-CBMAI, UNICAMP, Campinas, SP.Thermophilic fungus SSF was carried out using a 250 mL Erlenmeyer flask containing 5 g of sterilized mixture of sugar cane bagasse and orange bagasse in proportions of 30 and 70% (w/w). The material was inoculated with 5 mL of micelial suspension  which was obtained from a 4-day agar slant culture suspended in sterile distilled water. 4NO3, 0.1% NH4H2PO4, 0.1% MgSO4·7H2O at pH 5.0 was added to each of the several flasks. The final moisture content of the medium was approximately 70%. Cultivation was carried out at 45°C for 4 days. At 24 h intervals, the material corresponding to one Erlenmeyer flask was mixed with 40 mL distilled water, stirred for 40 min, filtered under vacuum, and centrifuged. The supernatant was used as crude enzyme solution.After inoculation, 10 mL of nutrient solution composed of 0.1% NHμmol of galacturonic acid per minute under the assay conditions.Exo-polygalacturonase (exo-PG) activity was assayed in a mixture containing 0.4 mL of 1% of citrus pectin solution (26% esterified—Sigma) in 0.2 M sodium acetate buffer (pH 5.5) and 0.1% of crude enzyme solution at 60°C for 10 min. The number of reducing groups, expressed as galacturonic acid released by enzymatic action, was quantified by the DNS method . One uniEndo-PG activity was measured viscosimetrically by adding 2 mL of crude enzyme to 6 mL of 0.2 M acetate-NaOH buffer (pH 5.5) containing 3% of low-esterified citrus pectin (Sigma). The reaction mixture was incubated at 60°C for 15 min, and its viscosity was determined with a basic viscosimeter (Fungilab). One unit of enzyme activity was defined as the amount of enzyme that reduced the initial viscosity by 50% per minute.150 mL of crude enzyme extract was dialyzed against 10 mM acetate buffer, pH 4.0, overnight. After dialysis, it was lyophilized and resuspended in 20 mL of 10 mM acetate buffer, pH 4.0.Gel filtration chromatography with Sephadex G-75 column (90.0 cm × 2.5 cm—Pharmacia) was used, and the elution occurred with 20 mM acetate buffer pH 4.0 at a flow rate of 16.8 mL/h. The PG activity (DNS assay method) and protein content of each tube were determined. The fractions containing the peak of enzyme activity were joined to the next step purification process in ion-exchange column.For ion-exchange chromatography, SP Sepharose column (20.0 cm × 2.5 cm—Aldrich) was used and the elution was made with same acetate buffer and flow at salt gradient from 0 to 1.1 M NaCl. The solution containing PG peak activity was desalted by dialysis against 10 mM acetate buffer, pH 4.0, at 4°C, overnight.The molecular weight of the purified enzyme was determined by SDS-PAGE in a Mini Protean II apparatus (10 × 8 cm) (Biorad). Electrophoresis was carried out in polyacrylamide gel, consisting of a 4% (w/v) stacking gel and 10% (w/v) resolving gel in Tris/glycine buffer (pH 8.3), by the method of Laemmli . The molAnalytical isoelectric focusing was performed in an Ettan IPGphor II Isoelectric Focusing system (Amersham) by electrophoresis in a 7.5% polyacrylamide gel (14 × 15 cm) containing 5% Pharmalyte (pH 3.0–10.0) (purchased from Amersham Bioscience). The gel was silver-stained to reveal protein.μg/mL by the microassay method of Bradford [Protein concentration was determined in the concentration of 10–100 Bradford , using bFor characterizing the PG activity, the DNS assay method was used. Optimal activity of purified PG was assayed as a function of pH, in 200 mM acetate buffer (pH 3.0–5.5), citrate-phosphate (pH 6.0–7.0), Tris-HCl (pH 7.5–8.5), and glycine-NaOH (pH 9.0–11.0), at 60°C with 2% low-esterified pectin (2%) as substrate. The pH stability of PG was evaluated by incubation of enzymatic solution in 0.1 M buffer solutions acetate (pH 3.0 to 5.5), citrate-phosphate (pH 6.0–7.0), Tris-HCl , and glycine-NaOH (pH 9.0–11.0), in the absence of pectin, for 24 hours. After this period, an aliquot was taken to measure residual activity under conditions of optimum pH and temperature. The effect of temperature on enzymatic activity was evaluated by incubation of reaction mixture at temperatures from 40°C to 80°C for 10 minutes, at optimum pH. The thermal stability was determined by measuring the residual activity of the enzyme after 1 h of incubation, in absence of substrate, at temperatures between 10 and 90°C. After this period, samples were taken to assay enzyme activity under conditions of optimum pH and temperature.To determine substrate specificity, solution of polygalacturonic acid, citrus pectin with 26% and 92% degree of esterification, and apple pectin (Sigma) were used as substrates at 2.0% in 0.2 M acetate buffer. The reaction was conducted in optimal conditions of enzyme activity.+3, Ag+, Ca+2, Mg+2, Mn+2, Zn+2, K+, and Hg+2) and EDTA at final concentration in the reaction medium of 2 mM. After 10 min. incubation at 4°C, the residual activity was measured under conditions of optimum enzyme activity.The influence of metallic ions on PG activity was evaluated by incubation of enzyme in the presence of different ionic solutions at 2 mM (FeKm) and Vmax values were determined from Lineweaver-Burk plots of enzyme activity measured with citrus pectin with 26% degree of esterification (Sigma) as substrates, at concentrations between 0.25 and 1.25% at optimum pH and temperature. The results were plotted with the program Grafit 5.0. The Michaelis constant , washed with alcoholic hydroxide silver for visualization of the bands. The mono-, di-, and trigalacturonic acids (Sigma) were used as standards. The crude enzyme solution obtained by fungus culture on solid-state fermentation applied on Sephadex G-75 gel column showed only one peak of enzyme activity, which was detected between 160.0 mL and 256.2 mL . This stIn the second step, 50 mL of enzymatic extract was applied on ion-exchange chromatography, using 20 mM acetate-NaOH buffer, at pH 4.0. Two protein peaks were observed from the elution volumes of 42.0 mL and 88.2 mL before the start of the salt gradient and three between 0.15 M and 0.7 M NaCl. Polygalacturonase was eluted at 0.9 M salt concentration . The speRhizopus oryzae and Niture and Pant [Fusarium moniliforme in solid-state fermentation.The samples application on gel electrophoresis indicated that the enzyme was purified to homogeneity and had molecular weight of 29.3 kDa , similarand Pant   were more resistant to pH and temperature changes compared to those obtained by submerged fermentation (SmF) [Thermomucor indicae-seudaticae showed higher thermostability in SmF than that in SSF.Pectinases obtained by on (SmF) . Moreoveon (SmF)  reportedIn relation to substrate preference, PG showed the highest activity with 26% esterified citrus pectin  similar + caused 18% decrease in PG activity  also inhibited about 50% enzyme activity of polygalacturonase produced by Fusarium oxysporum in SmF [Sporotrichum thermophile was inhibited by 78% with this ion at 1 mM [+2, similar results were found for other fungal PGs, which also suffered inhibition when exposed to this ion. The polygalacturonase produced by Thermomyces lanuginosus was inhibited by 53% [Sporotrichum thermophile was inhibited by around 50% with this ion at 1 mM [The ion Mgm in SmF . On the  at 1 mM . Regardid by 53% , while t at 1 mM .+2  [The PG activity was completely inhibited in the presence of Hg+2 , similar0.02 mM) . The eff0.02 mM) .Km of the PG was 1.58 mg/mL and Vmax was 1553.1 μmol/min/mg protein. The enzyme obtained from SmF showed Km of 0.62 mg/mL, and Vmax of 2433.2 μmol/min/mg [Km was high.The l/min/mg . These vKm of fungal polygalacturonases generally ranges from 0.12 to 6.7 mg/mL. These authors purified two polygalacturonases from Trichoderma reesei which had Km values of 0.15 mg/mL (PG I) and 0.93 mg/mL (PG II). Kaur et al. [Sporotrichum thermophile, with a Km of 0.416 mg/mL. The PG purified from the thermophilic fungus Acrophialophora nainiana showed a Km quite high (4.22 mg/mL), indicating low affinity for citrus pectin [According to Mohamed et al. , the Kmr et al.  purifieds pectin .The isoelectric point of the PG obtained from SSF was 6.6, a value different from that observed for enzyme produced in SmF which was 7.8 . These vT. aurantiacus are similar to those of other fungal strains described in the literature. García Maceira et al. [Fusarium oxysporum, which showed pI 7.0. Cabanne and Donèche [Botrytis cinerea. The authors observed that the enzyme from each strain showed different isoelectric points, ranging between 5.0 and 9.0.The pI values of the two PGs obtained by a et al.  purified Donèche  reported Donèche  describe Donèche , who repAfter incubation for 5 min at 65°C in 1% citrus pectin 26% DE methoxylation, the polygalacturonase released a mixture of mono-, di-, tri-, and oligogalacturonic acids  and was The PG activity measured by viscosimetric assay method (specific activity of 3542.8 U/mg) reduced afforded 56% of viscosity of 1% citrus pectin in 10 min. These results indicated an endo-PG activity. On the other hand, di and mono-galacturonates were released at the initial stages of the incubation period, suggesting that PG degraded the substrate by multiple attacks.Aspergillus kawachii in SmF. This PG was also unable to degrade digalacturonic and trigalacturonic acids but released unsaturated mono-, di-, tri-, and oligogalacturonic acids, indicating that it presents action in multiple attacks.This profile was very similar to that observed for the SmF enzyme by Martins et al. , which wThermoascus aurantiacus CBMAI756 in solid-state fermentation showed higher tolerance to variations of pH compared with PG produced in SmF by the same fungus, though that was less thermostable than SmF enzyme. Other enzymes features, such as isoelectric point, Km, and influence of some ions in the activity, were also different in relation to the fermentation process employed.The polygalacturonase obtained by thermophilic fungus


Aims. Although altered endocrine changes following bariatric surgery in morbidly obese patients with diabetes have been demonstrated by previous studies, little is known about their effects on low BMI patients of T2DM. We investigated the changes in adipokines and sICAM-1 in Chinese subjects with low BMI and T2DM after LRYGB and explored their relationship with postsurgical insulin sensitivity. Methods. Plasma levels of adiponectin, sICAM-1, fasting glucose, glycated hemoglobin, and fasting insulin and serum levels of visfatin were measured before and at three months after LRYGB in 33 T2DM patients with BMI of 22–30 kg·m−2. Results. Significant reductions in anthropometric measurements and indicators of glucose and lipid metabolism and moderate reductions in insulin resistance and fasting insulin were observed at three months after LRYGB. Postoperative adiponectin level (P < 0.001) was increased compared to the preoperative level, whereas visfatin (P < 0.001) and sICAM-1 (P < 0.001) were lower than that before surgery. Serum adiponectin negatively correlated with HOMA-IR and FIns both preoperatively and at three months after surgery, and visfatin positively correlated with HOMA-IR and FIns both preoperatively and postoperatively. Conclusion. Changes in adipokines were related to an improvement in postsurgical insulin sensitivity, which was predicted by weight loss after LRYGB even in low BMI patients with T2DM. The mean BMI of the postoperative participants whose adiponectin levels were below the median was significantly higher (25.81 ± 4.08 kg·m−2) than of those whose adiponectin levels were above the median (22.96 ± 2.38 kg·m−2). However, there was no significant correlation of these adipocyte hormones with BMI, WHR, FPG, 2HPG, HbA1c, TG, CHOL, HDL-CH, LDL-C, or sICAM-1. Serum sICAM-1 levels and changes in BMI, WHR, FPG, 2HPG, HbA1C, TG, CHOL, HDL-CH, LDL-C, HOMA-IR, FIns, adiponectin, and visfatin were not correlated.Concentrations of adipocyte hormones were further correlated with HOMA-IR and FIns. Both at baseline and at 3 months after surgery, adiponectin negatively correlated with HOMA-IR and Fins, whereas plasma visfatin positively correlated with HOMA-IR and FIns. A significant difference was found between the mean BMI of postoperative participants whose adiponectin values were below and above the median of adiponectin [−2 and 30.0 kg·m−2, respectively [−2 and increased atherogenic risk factors. A survey by Kim et al. reported that the prevalence of diabetes, hypertension, and dyslipidemia has doubled at a BMI of 23.0 to 24.0 kg·m−2 and tripled at a BMI of 26.0 kg·m−2 in the Korean adult population and that body fat and fat distribution at a BMI of 23.0 kg·m−2 in Asians may be similar to those in the Whites at a BMI of 25.0 kg·m−2 [−2) and obesity (BMI 25.0 kg·m−2) in Asian and Pacific Island populations [−2, and 63.6% patients showed T2DM remission, 27.3% showed glycemic control, and 9.1% showed improvement at 12-month duration in their study [−2 and that for obesity was 28 kg·m−2. In our study, 20 patients were overweight and 9 patients were obese. Some patients had raised triglyceride, cholesterol and/or LDL levels, and/or microalbuminuria. Global cutoff points for overweight and obesity as set by the World Health Organization are BMI of 25.0 kg·mectively . There hectively , 12, Japectively , Taiwan ectively , and Honectively  have rep0 kg·m−2 . The WHOulations . A studyir study . The BMI−2), and the mean age of the patients was (49.51 ± 1.33 years). The HbA1c level of all patients exceeded 7% despite being on maximum tolerated dose for a duration of at least six months. The major limitations of our study were that the sample size was limited to 33 individuals only, and as the sample was randomly selected, the patients could not be equally weighted for sex to allow for between sex differences. Expanding the sample size and the duration of study would further validate the results.Our study investigated the changes of adipocyte hormones and systemic marker of inflammation following LRYGB in low BMI Chinese subjects with T2DM. The mean BMI in our study was  were on oral hypoglycemic agents prior to surgery, and 8 (21.2%) were on insulin. The other 6 (33.3%) were using both oral hypoglycemic and insulin. After the surgery, 8 (24.2%) patients totally came off these medications, whereas 4 (12.1%) remained on insulin. The other 17 (51.5%) were on oral hypoglycemic agents but on a lesser dose than that before surgery.Three months after the surgery, statistically significant amelioration in body mass index and waist-hip circumference could be observed in our study. In addition, the indicators of glucose metabolism, namely, HbA1c, FPG, and 2HPG were significantly reduced in comparison to the preoperative values. Likewise, statistically significant reduction in TG could be seen. At three months post-LRYGB, decrease in HOMA-IR and FIns levels was also noted, suggesting that LRYGB could be beneficial even for low BMI individuals of T2DM . SimilarP < 0.001) three months after LRYGB  was noted   . Other sChanges in adipokines are related to an improvement in postsurgical insulin sensitivity even in low BMI patients with T2DM, and the major predictor was weight loss. Unraveling the pathogenesis and attaining in-depth exploration would require large-scale clinical studies and long-term followup that would further elucidate the effect of LRYGB in such patients.


Background. The Pakistani population has higher incidence of cardiovascular (CV) diseases at younger ages, due to undiagnosed, uncontrolled hypertension (HTN). A variety of associated HTN stressors is also reported. The study plans to understand the variables associated with initiation of HTN in this population. Objective. To find plasma aldosterone and cortisol relationship with some CV risk factors  in different stages of HTN particularly prehypertension. Subjects and Methods. The study conducted on 276 subjects (25–60 years), classified into prehypertensive (n = 55), HTN stage-1 (n = 70) and II (n = 76) according to 7th JNC report and compared with normotensive controls (n = 75). The anthropometric profiles  and BP recorded. Serum cortisol, aldosterone, total cholesterol, Low density lipoproteins, blood glucose, Na+ and K+, using standard laboratory techniques, were determined in fasting blood samples. Results. Subjects were mostly overweight and obese . The aldosterone level (ng/dl) was in higher normal range (9.17–12.41) and significantly correlated to BMI (0.587) in controls, and to TC (0.726) and LDL (0.620) in pre-HTN stage-I. The cortisol level was positively correlated (P < 0.01) to BMI (0.538), Na+ (0.690) and K+ (0.578) in control, and to BMI (0.628) and WC (0.679) in pre-HTN group, showing its association with BMI > 25. Conclusion. Pre-HTN stage among Pakistani population with successive increase in various risk factors of HTN in relation to aldosterone and cortisol has been identified. Interaction of the risk factors with endogenous levels of these hormones may initiate stages of HTN. Hypertension (HTN) is a major risk factor for target organ damage leading to many major diseases such as myocardial infarction, stroke, impaired renal function, and ultimately renal failure. Prevalence of HTN varies in different parts of the world, affecting 28% of adult population in North America, 40% in European countries, 25% in Far East region, 15% in South Asian countries, and 26% in Eastern Mediterranean region [Despite its high prevalence, identification of its key determinants remains challenging due to its multifactorial and polygenic nature. The genetic, environmental, and dietary factors possibly mediating via hormonal, metabolic, and neurological changes in tissues and organs are manifested as phenotypes ; howeverGenetic studies on Mendelian HTN have provided better understanding of etiological mechanisms including synthesis and degradation of mineralocorticoids and their receptors, renal sodium channels reabsorption mechanisms, regulation of kidney specific sodium-chloride cotransporters, and regulation of renin angiotensin aldosterone system. Thus, information about effects of adrenal hormones aldosterone and cortisol on myocardium, vascular endothelium and HTN is being accumulated . Aldoste+) or a fall in serum sodium (Na+) concentration. It exerts its effect initially by increasing epithelial Na+ channel (ENa+C) activity via an aldosterone specific enzyme eventually resulting in increased Na+ and decreased K+ levels [+ and water retention, accumulation of extracellular fluid volume, and increased cardiac output. This results in achievement of a steady state with renal K+ wasting and increase in arterial BP [The plasma aldosterone secretion is stimulated by either a rise in serum potassium  . The purCase-control study. Convenient.The study was approved by Board of Advanced Studies and Research, University of Karachi.n = 276) were classified into four groups according to cut-off values recommended by 7th JNC report [The subjects  representing urban population (as data was collected from largest city of country) were selected amongst patients attending five general practitioners clinics. Most of the subjects were educated and belonged to middle and lower middle socioeconomic class; details are published in previous article .Demographic data and lifestyle behaviors was collected by purpose designed questionnaire, followed by general physical examination.Written consent of every participant was taken.Subjects were weighed without shoes, in their normal clothing, using a digital scale with an accuracy of ±100 grams. Standing body height was measured without shoes to the nearest 0.5 cm by a commercial stadiometer with the shoulders in the relaxed position and arms hanging freely.2). The WHO recommended values for Asian population were taken as reference, >23 overweight and >25 obese [ Body mass index (BMI) was calculated from standard formula , total cholesterol (TC), and low density lipoproteins (LDL). Serum cortisol and aldosterone were measured by ELISA.Fasting venous blood samples were drawn (after 9–12 hours fasting), centrifuged, and analyzed  for estimation of fasting blood glucose (FBG), electrolytes  test was applied to compare pair-wise groups. Test of Pearson's correlation was applied to assess relationship of hormones with variables determined and shown by Scatter plots. Coefficient determination “n = 276) were almost equal with respect to sex . The mean age of control group was 37.5 ± 8.54, pre-HTN 39.2 ± 7.73, HTN stage 1 46.2 ± 12.1, and 47.5 ± 11.6 in stage II. Among healthy controls, 52 (69.3%) were found to be overweight (BMI > 23) with significantly higher proportions (P = 0.023) of overweight individuals in pre-HTN (80%), HTN stage I (90%),and stage II (76.3%) (The subjects ( (76.3%) .2) of control group (24.9 ± 3.77) was significantly less than that of pre-HTN group (26.4 ± 4.47), HTN stage I (28.4 ± 4.53), and stage II (26.6 ± 5.33) groups (P < 0.01). Mean WC of both HTN stage I (97.2 ± 11.4) and II (95.4 ± 13.3) was found to be significantly higher (P < 0.01) as compared to control (82.2 ± 13.4) and pre-HTN groups (86.9 ± 17.2) (The mean BMI (kg/m ± 17.2) . BMI has ± 17.2) .The mean systolic BP in various groups of control, pre-HTN, and HTN were 106.9 ± 10.7, 131.6 ± 7.1, 147.3 ± 8.4, and 171 ± 15.3. The diastolic BP were 85.9 ± 5.8, 93.7 ± 4.3, and 105.9 ± 9.7 respectively, as compared to control 68.4 ± 9.48 .P < 0.01), 171.7 ± 32.3 in pre-HTN (P < 0.05), and 179.1 ± 32.1 in stage II  level was higher in HTN stage 1(191.4 ± 48.4) as compared to 168.7±33.9 in controls (stage II . TC was stage II . The levstage II . LDL wasThe mean fasting blood glucose (FBG) level was in normal range in all groups but was highest in pre-HTN group (108.4 ± 38.2) as compared to controls (96.6 ± 38.4), HTN stage I (103.7 ± 32.7), and stage II (101.0 ± 43.6). FBG was correlated to LDL in pre-HTN group.The mean aldosterone level was the highest in, HTN stage I (12.41 ± 5.72) as compared to control group (9.17 ± 3.49), pre-HTN (8.76 ± 3.31), and stage II HTN (12.05 ± 6.84). The serum cortisol level in four groups was 7.61 ± 4.15, 9.36 ± 3.25, 9.08 ± 3.97, and 9.99 ± 4.85, respectively.+ concentration was within normal limits in all groups . Serum K+ level in four groups was also within normal range  showed that changes in BMI accounts for 39% of variation in cortisol and 48% interdependence between sodium and cortisol, whereas dependence shared between aldosterone and cholesterol was 53% and with fasting blood glucose 23% , dyslipidemia , and glycemic status of blood (FBG) in different stages of HTN. The study was focused on identification of risk factors in pre-HTN stage, that initiate an increase in set point BP, ultimately leading to progression of this stage into established HTN.Rapid urbanization and migration towards cities have brought significant changes in lifestyle patterns of our population, especially excessive dietary fat and salt intake and lack of physical activity, resulting in storage of surplus fat in subcutaneous and visceral adipose tissue  as evide+ retention, and HTN [+, and K+ in controls  converts cortisone to cortisol within insulin target tissue, such as liver, visceral adipose tissue and skeletal muscle, resulting in local GC action which by promoting adipogenesis  leads to and HTN . The mea converts Figures  and to B Figures . The relThe mean aldosterone level in this study was in high normal range and was found to have positive correlation with BMI and WC in controls, and to TC and LDL in pre-HTN group Figures  and 4. AA community-based study reported that even increased plasma aldosterone concentration within physiological range predisposed to the development of HTN . However+ and K+ are major determinants for the mechanical stiffness of endothelial cells. High plasma Na+ levels stiffen endothelial cells and block nitric oxide (NO) synthesis , whereas high plasma K+ levels soften endothelial cells and activate NO release [+ (10–16 mmol/day) coupled with usual Na+ intake (120–200 mmol/d) caused Na retention and an elevation of BP; on average systolic BP increased by 6 mmHg and diastolic BP by 4 mmHg in normotensive subjects and by 7 and 6 mmHg, respectively, in hypertensive subjects [Plasma Na release . Clinicasubjects .+ level was normal in all four groups; however, it was positively related to BMI and cortisol in controls and to systolic and diastolic BP in pre-HTN group. The strong relationship between Na+ and cortisol is very significant, as when in excess cortisol may act as an MC, perhaps by saturating the 11-beta hydroxysteroid-dehydrogenase-2 enzyme that inactivates cortisol at the renal tubules [+ metabolism at the cellular level may play a role in biochemical pathway leading to HTN [+ and a negative link of K+ to BP within a single population [Data analysis in this study indicated that Na tubules . A studyg to HTN  and anotpulation .+ level was within normal range and was positively related to cortisol in controls which is a significant finding, as high level of K+ is considered to be a stimulus for aldosterone secretion. Cortisol and aldosterone bind to the MC receptor with equal affinity, but normal circulatory concentration of cortisol is 100- to 1000-fold higher than those of aldosterone. If 11-beta hydroxysteroid (HSD2) is oversaturated or defective, more cortisol will be available to bind MCR [The serum Kbind MCR , 30, a cbind MCR .+ homeostasis is said to accompany many secondary forms as well as uncommon, inherited monogenic forms of HTN, as evidenced by 14% nonhypertensive subjects in Framingham heart study who developed HTN, when followed up to four years; however, other studies found no correlation between serum K+ and HTN [+ entry into the cells decreases, and its plasma level increases. The plasma K+ level need to increase only 1 meq/L to stimulate aldosterone secretion.Abnormal K and HTN . In IR, + status is unremarkable, cortisol appears to be responsible for MCR activation and aldosterone actions. Increased levels of these hormones in hypertensive subjects may be related to ACTH, which not only increases production of cortisol but also of aldosterone for short-term period [The visceral fat accumulation in upper abdomen also activates sympathetic nervous system and renin angiotensin system. When aldosterone levels are in low- to normal-range and Nam period . Individm period .+), in relation to aldosterone and cortisol in different stages of HTN. This study suggests that the interaction of these risk factors with endogenous levels of aldosterone and cortisol may disrupt the set-point BP and result in increased proportion of HTN. This study concludes the identification of pre-HTN stage among Pakistani population, with successive increase in various risk factors of HTN (obesity, dyslipidemia, hyperglycemia, and NaTimely and early detection of pre-HTN stage with a few preliminary investigations is an effective method of prevention of cardiovascular disease. The physicians are recommended to actively target lifestyle patterns for multiple risk reduction in these patients, as pre-HTN stage serves as an early warning sign for both patients and clinicians that metabolic changes which ultimately lead to CVD may well be underway. Moreover, aldosterone should be included in primary screening tests for evaluation of hypertensive patients.


The purpose of this review is to illustrate the wide spectrum of lesions in the corpus callosum, both congenital and acquired: developmental abnormalities, phakomatoses, neurometabolic disorders, demyelinating diseases, infection and inflammation, vascular lesions, neoplasms, traumatic and iatrogenic injury, and others. Cases include fetuses, children, and adults with rich iconography from the authors' own archive. Corpus callosum is one of the three interhemispheric commissures  and the greatest of them—it consists of approximately 190 000 000 axons . AnotherIn the anterior-posterior direction the corpus callosum is divided into rostrum, genu, body, isthmus, and splenium. The fibers in the rostrum connect fronto-basal cortex, in the genu—prefrontal cortex and anterior cingulate area, in the body—precentral (motor) cortex, insula, and cingulate gyri, in the isthmus—precentral and postcentral gyri  and primary auditory areas, and in the splenium—posterior parietal, medial occipital, and medial temporal cortexes .According to the newest theory the corpus callosum in its embryological development is fused of two separate parts: the anterior one, consisting of the rostrum, genu, and body and the posterior one—splenium. The place of the fusion is the isthmus. Callosal development is a very quick process and takes place in 13th week of gestational life. From this time on the corpus callosum grows mainly in the anterior direction, pushing the splenium posteriorly. It reaches its final shape in midgestation (week 20) but is still small and grows, initially by addition of fibers and later by myelination . The tarMyelination of the brain progresses from the center to the periphery, from bottom to top and from back to front. In newborns corpus callosum is not yet myelinated. In the 6th month after birth, when the cerebellum and genu of the internal capsule completed the process of myelination, the corpus callosum is myelinated in part (splenium), although it has not yet reached its target volume. Callosal genu is myelinated a little bit later than splenium—in about 8th month. It is not until about the first year of life that the corpus callosum displays the typical signal intensity: hyperintense on T1-weighted images and hypointense on T2-weighted images.There are a few papers illustrating callosal pathology in the literature –8. ContiAll the images were acquired with 1.5T scanners. Figures Philips Gyroscan ACS-NTSE/T1-weighted images (T1WI): repetition time TR shortest, echo time TE 14 ms, flip angle FA 90 deg, number of acquisitions NEX 2, matrix MX 256 × 225, field of view FOV 25 cm, slice thickness/interslice gap ST 5.0/1.0 mm,TSE/T2WI: TR shortest, TE 100 ms, FA 90 deg, NEX 2, MX 512 × 512, FOV 25 cm, ST 5.0/1.0 mm,FLAIR: TR 6000 ms, TE 100 ms, FA 90 deg, NEX 2, MX 256 × 256, FOV 25 cm, ST 5.0/1.0 mm,FFE/T2∗WI: TR shortest, TE 23 ms, FA 15 deg, NEX 2, MX 256 × 256, FOV 23 cm, ST 5.0/1.0 mm,DWI: TR shortest, TE 42 ms, FA 90, NEX 1, ST 6.0/1.0 mm.GE Signa HDxt (adult brain)SE/T1WI: TR 540 ms, TE 9 ms, NEX 2, MX 320 × 224, FOV 24 × 18 cm, ST 5.0/1.5 mm,FSE/T2WI: TR 5000 ms, TE 88 ms, NEX 1.5, MX 384 × 384, FOV 24 × 24 cm, ST 5.0/1.5 mm,FLAIR: TR 8002 ms, TE 126.8 ms, TI 2000, NEX 1, MX 320 × 256, FOV 24 × 24 cm, ST 5.0/1.5 mm,GRE/T2∗WI: TR 660 ms, TE 15 ms, NEX 2, MX 320 × 192, FOV 24 × 18 cm, ST 5.0/1.5 mm,b = 1000.DWI: TR 8000 ms, TE 98.5 ms, NEX 1, MX 128 × 128, ST 5.0/0.0 mm, GE Signa HDxt (children)SE/T1WI: TR 540 ms, TE 9 ms, NEX 2, MX 320 × 224, FOV 24 × 18 cm, ST 5.0/1.5 mm,FSE/T2WI: TR 5500 ms, TE 84 ms, NEX 1.5, MX 320 × 320, FOV 24 × 24 cm, ST 5.0/1.5 mm,FLAIR: TR 8000 ms, TE 140.2 ms, TI 2000, NEX 1, MX 320 × 320, FOV 24 × 18 cm, ST 5.0/1.5 mm,GRE/T2∗WI: TR 660 ms, TE 15 ms, NEX 2, MX 320 × 192, FOV 24 × 18 cm, ST 5.0/1.5 mm,SWI: TR 6750 ms, TE 40 ms, NEX 4, MX 256 × 512, FOV 26 × 26 cm, ST 3.0/0.0 mm,b = 1000,DWI: TR 6625 ms, TE 100.5 ms, NEX 2, MX 160 × 160, ST 5.0/1.5 mm, FSPGR/3D/T1WI: TR 8.1 ms, TE 3.6 ms, TI 450, NEX 1, MX 320 × 224, FOV 24 × 24 cm, ST 1.6/−0.8 mm,CUBE/3D/FLAIR: TR 6000 ms, TE 130.7 ms, TI 1852, NEX 1, MX 224 × 224, FOV 22 × 22 cm, ST 1.4/−0.7 mm.The following acquisition parameters were used:Incomplete or abnormal development leads to the most common pathology, which affects the organs in question: agenesis and hypoplasia.The characteristic appearance of callosal agenesis makes this anomaly easily and early recognizable on prenatal ultrasound and MRI: wide interhemispheric fissure , upward MRI allows for visualization of the bundles of Probst—evidence that callosal fibers are not really agenetic but heterotopic, lying parasagittally on both sides and giving the lateral verticle appearance of moose head or viking helmet on coronal images.In contrast to patients after callosotomy, individuals with callosal agenesis show only weak evidence of a “disconnection syndrome” which suggests that brain plasticity allows for forming alternative pathways of interhemispheric transfer in cases of this congenital anomaly .One has to remember that as the three interhemispheric commissures develop together, callosal agenesis is only rarely isolated: it is accompanied by hippocampal commissure agenesis and in 50% of cases also by anterior commissure agenesis or hypoplasia ..12].Phakomatoses belong to congenital diseases in which callosal abnormalities are observed. Neurofibromatosis type 1 or von Recklinghausen disease is the most frequent of them, with the estimated incidence of 1 : 3000. Neurofibromatosis bright objects (UNO), called formerly unidentified bright objects (UBO), are T2 hyperintense and appear most often in the basal ganglia, brainstem, and posterior fossa. They are also found in the corpus callosum, mainly in the splenium Figures . UNO areHigher incidence of callosal agenesis/dysgenesis is described in other neurocutaneous syndromes, among them are the Sturge-Weber syndrome, tuberous sclerosis complex, and Bloch-Sulzberger syndrome  15]..15].X-ALD is an inborn disorder of peroxisomal fatty acid beta oxidation which results in the accumulation of very long chain fatty acids in tissues. It affects mainly the myelin in the central nervous system, the adrenal cortex, and the Leydig cells in the testes. In the most typical form which accounts for approximately 80% of the cases demyelination involves callosal splenium and spreads symmetrically into occipital and parietal lobes  and thenThe anterior part of the corpus callosum is involved in the Alexander disease. Callosal demyelination is observed in many neurometabolic diseases, for example, in globoid leukodystrophy (Krabbe disease) , metachrIn the course of neurometabolic diseases callosal agenesis is also observed, among others in nonketotic hyperglycinemia, Menkes kinky hair disease, Hurler disease or maple syrup urine disease . In otheCallosal involvement is typical of MS although it has never been included in the evolving diagnostic criteria of this disease . The typThe Marchiafava-Bignami disease is characterized by callosal demyelination and necrosis with subsequent atrophy. It is classically associated with chronic alcoholism but it has also been described in patients with malignancy and nutritional deficiencies. The lesions are T2- and FLAIR hyperintense which reflects edema and myelin damage. Necrosis in the chronic stage is reflected by T1-hypointensity but lesions may also regress .Cerebrovascular involvement is common in group B streptococcus meningitis, especially in neonates, but also in older children. There are two main patterns of brain infarction: deep perforator arterial stroke to basal ganglia, thalamus, and periventricular white matter and focal cortical infarctions . In our Lyme disease, caused by Borrelia burgdorferi, belongs to infectious diseases that are most commonly mistaken for MS ..25].Corpus callosum has rich blood supply from the anterior communicating artery , the pericallosal artery which supplies the body, and the posterior pericallosal artery, a branch of the posterior cerebral artery, which feeds the splenium. Isolated callosal infarcts are therefore uncommon. If present, they affect callosal splenium more often than the body and genu  26]. Th. Th26]. Arteriovenous malformations of the corpus callosum account for 9–11% of all cerebral AVMs . They arGlioblastoma multiforme (GBM) (WHO grade IV) is the most common and most aggressive malignant primary brain tumor. Callosal GBM, in addition to the corpus callosum, affects also both cerebral hemispheres resulting in the typical “butterfly glioma” appearance with solid intense contrast enhancement in the corpus callosum .Gliomatosis cerebri, WHO grade III, does not form a solid tumor but diffusely infiltrates the brain tissue. The architecture of the brain is preserved but the affected portions of the brain are swollen. Loss of distinction between grey and white matter is observed. Usually bilateral widespread invasion with involvement of the corpus callosum is found Figures  28]. In. In28]. Oligoastrocytoma (mixed glioma) occurs in two main types: well-differentiated oligodendroglioma (WHO grade II) and its anaplastic variant (WHO grade III). The most frequent locations are the frontal lobes and these tumors may involve the corpus callosum and extend through it to the contralateral hemisphere producing a “butterfly glioma” pattern Figures . Signal Primary CNS lymphoma accounts for approximately 16% of primary brain tumors. Most of them are non-Hodgkin's and represent B-cell type. They are most often isointense-hypointense on T1-weighted images, hypointense on T2-weighted images and enhance homogeneously with gadolinium-based contrast media. In classic presentation the tumors involve the corpus callosum in a butterfly pattern . In patiCorpus callosum may also be affected by metastases although it is reported to be rare. Callosal involvement is more frequent in case of infiltration by a lesion from the adjacent structures . In our The term “diffuse axonal injury” (DAI) refers to extensive traumatic lesions in white matter tracts. This kind of injury is the result of traumatic shearing forces that occur when the head is rapidly accelerated, decelerated, or rotated. Motor vehicle accidents are the most frequent cause of DAI but it can also be a result of child abuse, for example, in shaken baby syndrome. Corpus callosum belongs to the most frequently injured parts of white matter. The splenium and the undersurface of the posterior body are mainly involved due to vicinity of the falx cerebri. The lesions are typically small (1–15 mm) and invisible on CT. MRI is a method of choice in their diagnosis. They are T2-hyperintense but first of all show diffusion restriction with reduced ADC values  32, 33], 3332, 3 After hemorrhagic injury hemosiderin deposits may be seen in the corpus callosum . It may Corpus callosum may be injured as a result of shunting procedures in patients with hydrocephalus Figures  and 31 .Posterior reversible encephalopathy syndrome (PRES) is a toxic-metabolic disease characterized by headache, confusion, seizures, and visual loss. It occurs in patients with malignant hypertension, eclampsia, hypercalcemia, receiving some drugs, for example, cyclosporine, after organ transplantation. That is why the condition may be considered as iatrogenic. The brain swelling is seen on MRI mainly in the posterior parts of the brain, including splenium of the corpus callosum. The symptoms tend to resolve after a period of time  35]..35].Perivascular Virchow-Robin spaces may be seen in the corpus callosum as an incidental finding . AbnormaCallosal atrophy is associated with aging . In Alzheimer's disease (AD) callosal atrophy reflects loss of intracortical projecting neocortical pyramidal neurons and is more severe than in healthy subjects. The most significant atrophy in AD is noticed in callosal splenium. Callosal atrophy correlates with progression of dementia severity in AD patients .Linear T2- and FLAIR hyperintensity of the ventral part of the corpus callosum is a frequent finding reflecting gliosis and is attributed in the literature to the elderly age, subcortical arteriosclerotic encephalopathy, and radiation therapy , 37. In Transient splenial lesion is a term attributed to the ovoid or round focus in the central part of the callosal splenium that has been described in cases of epilepsy and encephalitis. These lesions show diffusion restriction and regress with time they are regarded as intracellular (intramyelinic) edema . In our Being the largest brain commissure, the corpus callosum is related to cognitive functions, social skills, problem solving, and attention. Thanks to its multiplanar nature and high tissue resolution magnetic resonance imaging is a method of choice in the assessment of the corpus callosum and its congenital and acquired pathological lesions. It is a perfect diagnosing tool from the very beginning of life, that is, from the prenatal period. Visualization of callosal involvement helps to establish diagnosis in certain disease entities.


Introduction. Lactobacillus casei and Lactobacillus acidophilus were used to assess the anti-inflammatory properties in carrageenan induced acute inflammatory model. Materials and Methods. Diclofenac sodium was used as standard drug at concentration of 150 mg/kg of body weight. Culture of Lactobacillus  2 × 107 CFU/ml was given orally. Edema was induced with 1% carrageenan to all the groups after one hour of the oral treatments. Paw thickness was checked at t = 1, 2, 3, 4, 5, and 24 hours. Stair climbing score and motility score were assessed at t = 24 hours. Cytokines assay for IL-6, IL-10, and TNF-α was performed on serum samples. Results. Lactobacillus showed a statistically significant decrease in paw thickness at P < 0.001. L. acidophilus and L. casei decreased by 32% and 28% in paw thickness. They both significantly increased the stair climbing and motility score. Lactobacillus treatment significantly downregulated IL-6 and TNF-α while upregulated IL-10 at P < 0.0001. Conclusion. L. casei and L. acidophilus significantly decreased the inflammatory reactions induced by carrageenan. This study has also proposed that Lactobacillus ameliorated the inflammatory reaction by downregulating the proinflammatory cytokines pathway. Inflammation is a local response of living mammalian tissues to injury. It is a body defense reaction in order to eliminate or limit the spread of injurious agent . There aLactobacillus and its important species like L. casei and L. acidophilus have been used against many pathological and disease conditions. Lactobacillus is known for its antimicrobial, antilipidemic, immunomodulatory, anticancerous, antidiabetic, and antiarthritic properties [Lactobacillus has been assessed for its immunomodulatory properties in different experiments [ As it is well known, the probiotic bacteria are nonpathogenic and consumed as/with food since a long time. The use of probiotic bacteria in dietary supplements or dietary products is widely documented in the literature. operties –10. Lacteriments –15.According to the WHO report, about 70–80% of the world's population rely on nonconventional medicine mainly from herbal sources in their primary health care , 17. EspL. casei and L. acidophilus using carrageenan-induced acute inflammatory model. With these facts taken into account, present study was planned to find out the possibility of anti-inflammatory activity of Lactobacillus acidophilus (ATCC 314) and Lactobacillus casei (ATCC 334) were purchased from Hi Media, Navi Mumbai, India. Lyophilized culture was streaked over de Mann Rogosa Sharpe Agar (MRS) at 37°C in anaerobic condition.Carrageenan was purchased from Hi Media, Mumbai, India. Standard anti-inflammatory drug diclofenac sodium was purchased from Recon, Bangalore, India. Cytokines assay kits were purchased from Ray Biotech, Norcross GA, USA and DNA Bio, Hyderabad, Andhra Pradesh, India.ad libitum. All animals were acclimatized for at least one week before the experimental session. All the experimental procedures were done following the guidelines of the Institutional Animals Ethics Committee (IEAC).35 male Wistar rats (200 gm each) were used for the present study. They were fed with standard pellet diet and water For the anti-inflammatory activity against the acute inflammation, animals were divided into five groups.μL of distilled water; Group C and Group D received 2 × 107 CFU/mL of L. casei and L. acidophilus, respectively, suspended in 500 μL of distilled water. Group E (positive control) animals were administered with diclofenac sodium (150 mg/Kg body weight).Group A (carrageenan control) did not receive any oral treatment; Group B (control) received 500 μL of 1% freshly prepared solution of carrageenan in distilled water into the right-hind paws of each rat of all the groups except the group A. Animals of group B/C, D/E were treated with the single dose of vehicle, cultures, and drug, respectively; 30 minutes prior to carrageenan injection. Paw thickness were measured just before the carrageenan injection, that is, at “0 hour” and then at 1, 2, 3, 4, and 24th hour after carrageenan injection. Increase in paw thickness was measured as the difference in paw thickness at “0 hour” and paw thickness at respective hours.Anti-inflammatory activity was measured using carrageenan-induced rat paw edema assay , 21. EdeOvernight fasting animals were trained for one week to climb a staircase with steps at 5, 10, and 15 cm having water at the second and food at the third step. Climbing ability of the rats in above groups was scored 0 if the rats did not climb; 1, if the rats climbed onto step 1; 2, if the rats climbed onto step 2; 3, if the rat could climb all the three steps , 23.The motility pattern of the rats was observed for a period of 5 minutes and scored 0, if the rat walked with difficulty and avoided touching the toes of the inflamed paw to the floor; 1, if the rat walked with little difficulty, but with toe touching the floor; 2, if the rat walked easily , 23.After 24 hours, all animals were sacrificed with cardiovascular bleeding with the help of Di ethyl ether according to the guidelines of CPSCEA committee. Blood was collected for the cytokines assay. Blood samples were left to coagulate for room temperature for 60 minutes and then centrifuged at 1500 g for 15 minutes and crude serum was kept in new tubes. Serum was harvested and stored at −20°C until use.α (proinflammatory cytokines) in picogram per millilitre (pg/mL) were estimated with the help of ELISA Reader . Serum samples were used. IL-6 and TNF-α (Ray Bio ) and IL-10 (DNA bio) ELISA kits were used. Assays were performed according to the manufacturer's recommendations.IL-10 (anti-inflammatory cytokines) and IL-6, TNF-± SEM of seven observations and ANOVA followed by post hoc test. Duncans test was used to compare the groups. The stair climbing ability test and motility are expressed as median scores and the Kruskal-Wallis test was used to compare the groups.The value for edema volume is expressed as mean t = 0 was 3.023 ± 0.0408 cm and this remains constant at the end of 24 hours. Group B animals had showed an increase in paw thickness at each hour which was significant at P < 0.001. At 0 hours the thickness was 3.028 ± 0.040 cm, which increased to 3.59 ± 0.049 cm at t = 3 hours. At 24 hours the thickness was found to be 4.01 ± 0.025 cm. The paw thickness of Group C animals was 3 ± 0.028 cm which showed a mild increase at the end of 2nd hour, that is, 3.35 ± 0.0102 cm. After the 2nd hour it decreased to 3.04 ± 0.077 cm, 3.014 ± 0.0489 cm, 2.99 ± 0.0053 cm, and 3.014 ± 0.024 cm at the end of 3, 4, 5, and 24 hours, respectively. Group D animals showed an increase up to the 2nd hour. 3.03 ± 0.065 cm thickness was found at the end of third hour, which decreased to 3.0014 ± 0.0024 cm at t = 24 hours. So, Groups C and D indicated a statistically significant decrease in paw thickness (P < 0.001). Group E animals also showed an increase in paw thickness of 3.042 ± 0.077 cm (t = 0 hours), 3.26 ± 0.069 cm (t = 1 hours), 3.22 ± 0.0184 cm (t = 2 hours), and 3.074 ± 0.0762 cm (t = 3 hours). It increased after the third hour to 3.174 ± 0.057 cm (t = 24 hours). These values were found to be statistically significant at P < 0.001 . Lactobacillus treatment significantly increased the stair climbing score of 2.57 ± 0.489 and 2.71 ± 0.408 for Group C and Group D respectively (significant at P < 0.001). Group E animals were showing a score of 1.71 ± 0.6122. Group B animal score was lowest in comparison to Groups A, C, D, and E animals . Score for Group A animals was  2 ± 0.00  whereas 0.14 ± 0.244 was found in case of group B animals. This was found to be the lowest when comparing Group E and Group B animals . On the contrary, Lactobacillus treatment statistically decreased the IL-6 and TNF-α in Group C and Group D at P < 0.0001. Lowest value for IL-6 was observed in case of Group D at 44.505 ± 0.198 pg/mL, while lowest value of TNF-α was observed for Group C at 420.77 ± 0.265 pg/mL. Group E animals was showing IL-6, TNF-α, and IL-10 values of 64.60 ± 0.131 pg/mL, 490.52 ± 0.228 pg/mL, and 29.675 ± 0.075 pg/mL, respectively. Only IL-6 concentration for Group E animals was not significant at P < 0.0001 while IL-10 and TNF-α concentrations were significant at P < 0.0001  [Carrageenan-induced rat paw edema model is a suitable test for evaluating anti-inflammatory drugs, which has frequently been used to assess the antiedematous effect of the drug. Carrageenan is a strong chemical use for the release of inflammatory and proinflammatory mediators  .Lactobacillus sp might be containing some anti-inflammatory agent which is responsible for the blockage of prostaglandins and inflammatory pathway. The course of acute inflammation is biphasic. First phase starts with the release of histamine, serotonin, and kinins after the injection of phlogistic agent in the first few hours . While tL. casei and L. acidophilus had very consistent anti-inflammatory activity and thus showed significant decrease in the paw thickness of rat (Group C and Group D). Previously, some researchers have also proposed the anti-inflammatory property of Lactobacillus in gut [In this model of inflammation, s in gut –29. Alths in gut .Lactobacillus might have inhibited the cyclooxygenase that synthesizes prostaglandins. Oral administration of Lactobacillus significantly downregulated the proinflammatory cytokines while upregulated the anti-inflammatory cytokines. Prostaglandins play essential role in inflammation [L. casei and L. acidophilus overcome the inflammation induced by carrageenan. Our present findings are consistent with the results of some other workers [α, IL-1, IL-6, IL-8, and GM-CSF by human monocytes [ammation . Prostag workers , 31–34. onocytes .γ and TNF-α release [Knowledge on the immune-mediated mechanisms in metabolic scenario has markedly increased in the recent past, evidencing the role that dietary components may have to modulate immunity by enhancing or suppressing the immune response. For instance, certain strains of probiotics have been demonstrated to be able to modulate the immune system by stimulating release of different patterns of cytokines by different cells. It could be able to modify the number of CD4 T cells and actively interfere with anti-inflammatory and proinflammatory signalling pathways by inducing production of IL-10 and reducing INF- release . ProbiotLactobacillus suppresses the first phase of carrageenan-induced paw edema, thus, confirming an NSAID-like property. The present study showed that L. casei and L. acidophilus have both analgesic and anti-inflammatory properties.The present results suggest that