Mel decreased cytolysis and lipid peroxidation and improved renal function and morphology compared to I/R group. Parallely, it significantly reduced the ER stress parameters including GRP 78, p-PERK, XBP 1, ATF 6, CHOP, and JNK. Simultaneously, p-Akt level was significantly enhanced and its target molecules GSK-3β and VDAC were inhibited. Furthermore, the ERK and P38 phosphorylation were evidently augmented after Mel administration in comparison to I/R group. In conclusion, Mel improves the recovery of renal function by decreasing ER stress and stimulating Akt pathway after renal I/R injury.Melatonin (Mel) is widely used to attenuate ischemia/reperfusion (I/R) injury in several organs. Nevertheless, the underlying mechanisms remain unclear. This study was conducted to explore the effect of Mel on endoplasmic reticulum (ER) stress, Akt and MAPK cascades after renal warm I/R. Eighteen Wistar rats were randomized into three groups: Sham, I/R, and Mel + I/R. The ischemia period was 60 min followed by 120 min of reperfusion. Mel (10 mg/kg) was administrated 30 min prior to ischemia. The creatinine clearance, MDA, LDH levels, and histopathological changes were evaluated. In addition, Western blot was performed to study ER stress and its downstream apoptosis as well as phosphorylation of Akt, GSK-3 Ischemia/reperfusion (I/R) injury remains a major problem encountered in vascular surgery, organ procurement, and transplantation and can lead to structural and functional cell damage . In renaER plays an important role in synthesis and maturation of proteins, biosynthesis of lipids, regulation of calcium, and maintenance of cell homeostasis . Disturbβ pathway.Melatonin (N-acetyl-5-methoxytryptamine) (Mel) is a hormone secreted by pineal gland and is mainly responsible for controlling circadian cycle . It is aMale Wistar rats weighing 200–250 g were used in this study. Animals were maintained at constant temperature (23 ± 2°C) with a 12 h light-dark cycle and free access to water and food. All procedures were carried out in accordance with the European Union Regulations (Directive 86/609/CEE) for animal experiments. Animals were randomly assigned into the following experimental groups, each containing 6 rats.I/R Group. Rats had been injected with a vehicle solution consisting of ethanol and NaCl 0.9% mixture  intraperitoneally (i.p.) 30 min before they were subjected to bilateral renal ischemia. The renal pedicles were occluded for 60 min using nontraumatic vascular clips, followed by reperfusion for 120 min [ 120 min . Mel + I/R Group. This group was the same as I/R group but animals were treated with Mel (10 mg/kg i.p.) 30 min before renal clamping [clamping , 14. MelSham Group. Animals were subjected to the same surgical procedure described above but were not subjected to renal I/R and did not receive any treatment.As described previously by Mahfoudh-Boussaid et al.  rats werAt the end of the reperfusion period, rats were euthanized and blood samples were collected via carotid artery. Simultaneously, both kidneys were harvested and weighed. Plasma, urine, and tissue samples were immediately kept at −80°C for biochemical and Western blot analyses.μmol/L), V is urine flow rate (μL/min/g of weight), and Creat P is the creatinine concentration in plasma (μmol/L).The renal function was evaluated during the last thirty minutes of reperfusion by calculating the creatinine clearance using the standard formula:Serum and urine creatinine concentrations were measured spectrophotometrically at 490 nm by the Jaffé kinetic reaction .The activity of lactate dehydrogenase (LDH) in the plasma was quantified using LDH assay kit .Malondialdehyde (MDA) is an end product of peroxidation of cell membrane lipids caused by oxygen derived free radicals. It was measured in renal tissue using the thiobarbiturate reaction at 530 nm .Samples from all kidneys were fixed in 10% formalin solution and processed for histology by hematoxylin and eosin (H&E) staining. Histological evaluations were performed using light microscopy at a magnification of 40–400 by an experienced renal pathologist without having knowledge about the treatment groups. Morphological changes from the whole cross-sectional area of the kidneys were assessed and a semiquantitative analysis of histological damage was conducted. A score from 0 to 4 was given to assess necrosis according to the method of Jablonski et al.  as the fμg/lane) were then separated by SDS-PAGE electrophoresis and transferred to polyvinylidene fluoride membranes. Membranes were immunoblotted with antibodies directed against GRP78, total and p-PERK, XBP-1, ATF6α and CHOP , β-actin , total and p-JNK, total and p-Akt, total and p-GSK-3β, total and p-ERK, total and p-P38 , and total and p-VDAC. The bands were visualized using an enhanced chemiluminescence kit . The values were obtained by densitometric scanning and the Quantity One software program .Western blot analysis was performed as previously described by Mahfoudh-Boussaid et al. . Brieflyp < 0.05 was considered statistically significant.The data are presented as means ± SEM. Statistical analysis was performed using one-way ANOVA, followed by Newman-Keuls multiple comparisons. p < 0.05). This rise was significantly lower after Mel treatment with values of 930 ± 69 U/L compared to I/R group (p < 0.05).Cytolysis was assessed by measuring the activity of LDH in the plasma . Level op < 0.05). After the administration of Mel, we observed a significant drop in this parameter reaching 0.21 ± 0.02 nmol/mg protein regarding I/R group (p < 0.05).Then, we examined the protective effect of Mel on lipid peroxidation . Renal IμL/min/g of weight compared to those of Sham group with values of 3.8 ± 0.4 μL/min/g of weight (p < 0.05). An important functional recovery was observed subsequently to Mel administration prior to renal I/R and the creatinine clearance was 3.36 ± 0.3 μL/min/g of weight (p < 0.05) in comparison to I/R group.Renal function was evaluated comparing the variation of creatinine clearance among the three groups . Kidneysp < 0.05). In comparison to the I/R group, treatment with Mel significantly attenuated this score reaching 1.83 ± 0.2 (p < 0.05).Histological changes were in keeping with biochemical parameters of renal injury. The morphological examination of all groups confirmed that there was renal impairment by severe tubular damage after I/R  comparedα from Sham values of 98 ± 23, 56 ± 20, 62 ± 4, and 73 ± 61, respectively, to 391 ± 8, 205 ± 24, 305 ± 51, and 160 ± 21, respectively (p < 0.05). Interestingly, rats undergoing Mel treatment demonstrated a lower level activation of these ER stress parameters in the kidney compared with their respective I/R groups with values of 227 ± 2, 105 ± 13, 102 ± 22, and 66 ± 5, respectively (p < 0.05).Afterwards, we examined the possible involvement of Mel in modulating ER stress induced by I/R injury. Different ER stress pathways were explored . Renal Ip < 0.05). In contrast, this effect was significantly attenuated by preischemic Mel administration and values dropped to 123 ± 13 and 267 ± 4, respectively, in comparison to I/R solely.In line with this, we studied the ER stress induced apoptosis by focusing on two proapoptotic parameters. As indicated in p < 0.05) (p < 0.05) (β phosphorylation (p < 0.05). Moreover, GSK-3β phosphorylation was significantly enhanced after Mel treatment to 354 ± 36 in comparison to both other groups.Furthermore, we evaluated the influence of Mel on some targets of the Akt signaling pathway. We found that Akt phosphorylation was notably decreased after I/R to values of 93 ± 4 when compared to Sham where p-Akt level was 141 ± 18 ( < 0.05) . This wa < 0.05) . Howeverrylation . The admp < 0.05), in comparison to both other groups.To finally explore the relevance of Mel treatment on some MAPK features after exposure to renal I/R, immunoblot analyses of ERK and P38 in kidneys from all groups were performed. As shown in The effect of Mel without I/R (Sham + Mel group) was assessed regarding the parameters used in the present study (data not shown) and no significant differences were recorded in comparison with Sham group. Therefore, only the data obtained in Sham group were mentioned in this study and were used for further statistical analysis with I/R and Mel + I/R groups.Renal I/R injury remains an unresolved problem that has immediate and deleterious effects in both native and transplanted kidneys . The patIn addition to the decrease of I/R induced elevation in lipid peroxidation, our findings revealed that Mel reduced LDH release and improved the creatinine clearance of ischemic kidneys. It has been suggested that lipid peroxidation is closely related to I/R induced tissue damage and that MDA is an indicator of lipid peroxidation rate . The perThe next finding of this study shows that Mel significantly attenuated the level of several ER stress parameters induced by renal I/R. It is well-documented that ER stress plays a significant role in the pathogenesis of renal I/R injury , 29. In β and VDAC. Results of the current study showed that p-Akt decreased in the case of I/R injury and that Mel prevented this downregulation. Many researchers have demonstrated that Akt plays a crucial role in the protection of liver [β which decreases the level of phosphorylated VDAC, the most abundant protein in mitochondrial outer membrane, leading to inhibition of the apoptotic process [To further elucidate the mechanisms by which Mel modulates cell survival during renal I/R, we evaluated its effects on Akt and its downstream targets, GSK-3of liver , heart [of liver  and kidnof liver  against  process , 42. The process  and live process  but is nβ activity [There are likely many other mechanisms by which Mel can promote cell survival. In order to explore other possible pathways, we examined the phosphorylation levels of ERK and P38 which are serine/threonine kinases belonging to the MAPK family. ERK activation is commonly considered as protective in the setting of renal I/R . Besidesactivity  thus conactivity . P38 is activity . Accordiactivity . Other ractivity . Importaactivity . Hernendactivity . It is oOn the basis of the previous findings, we conclude that Mel could ameliorate renal damage related to I/R. The underlying mechanisms of this protection essentially involve the modulation of ER stress, Akt and MAPK pathways.


P < 0.05) and performance (P < 0.05). However, heart rate and SJFT index (P < 0.05) increase significantly during CR in comparison to baseline. Moreover, exercise leads to a significant increase in testosterone, cortisol, growth hormone (GH), leukocytes, neutrophils, TNF-α, and IL-6, in both CR and baseline conditions. Compared to baseline, TNF-α and IL-6 were significantly higher during CR condition (P < 0.05). Additionally, CR leads to an increase in cortisol and GH (P < 0.05) and a decrease in testosterone concentrations (P < 0.05).The aim of this study was to evaluate the effect of caloric restriction on the immune and hormonal responses during exercise in judo athletes. In a randomised order, 11 male judokas  participate in this study during a period of weight maintenance (baseline) and after 7 days of caloric restriction (CR). All subjects performed the Special Judo Fitness Test (SJFT) during the two conditions. Values for nutrient intakes were obtained from a 7 d food record kept during a period of weight maintenance and after a 7-day food restriction (−5~6 MJ/day). Our results showed that CR resulted in significant decreases in body weight ( In judo competition, division by weight class guarantees matched strength, agility, and power between the competitors , 2. To bOn the other hand, pervious reports showed that strenuous high-intensity exercise induces immune suppression and may explain the increased risk of infection in athletes , 12. LikIt is also well established that restriction of energy and nutrients results in a compromised immune system and, hence, a decreased resistance to infection. Likewise, CR appears to synchronize the central pacemaker in the SCN, suggesting a role for a metabolic state imposed by low calories in central clock entrainment. There is a general perception among athletes, coaches, and team physicians that athletes are more vulnerable to infections disease. Moreover, rapid weight reduction by energy restriction may have additional disadvantages in athletes who are in a chronic immunosuppressive state. In this context, Shimizu et al.  showed tResults of several studies have suggested adverse effects of weight loss, including intensive training and energy restriction, on immune parameters in athletes. Weight loss reportedly reduces neutrophil phagocytic activity , T-cell Additionally, studies investigating the effects of weight loss on performance utilized laboratory-based techniques, which may not reflect the demands of real judo combat. Therefore, a judo-specific performance test , which is more representative of judo movements than laboratory tests, has been proposed as a valid and reliable measure of performance in judo athletes , 23. Howα, testosterone, and cortisol during SJFT in judo athletes. In light of the literature observations, we hypothesized that CR affects the proinflammatory cytokines and steroid hormone responses to the SJFT.It is crucial, therefore, to investigate the effect of 7 days' food restriction on proinflammatory cytokines and steroid hormones in male judo athletes preparing for a national championship. In view of the above consideration, the purpose of this study was to determine the effects of caloric restriction on IL-6, TNF-Eleven healthy judo athletes (mean (±SD); age: 20.45 ± 2.51 years; height: 171.6 ± 3.6 cm; and weight 75.9 ± 3.1 kg) participated in the study. After receiving a thorough explanation of the protocol, they gave written consent to participate in this study. This study protocol was in accordance with the Helsinki Declaration for human experimentation and was approved by the university ethics committee. The participants were also selected based on their chronotype and on the basis of their answers to Horne and Ostberg  self-assDuring the week before the experiment, participants came to the laboratory several times to become fully familiarized with the procedure and tests involved so as to minimize learning effects during the experiment. In a randomized order, subjects participated in two experimental test sessions. The first was a baseline condition in which subjects were taking a normal diet (baseline). The second is a condition of caloric restriction for 7 days; participants reduce their energy intake by 6.7 MJ/day (CR). During each experimental condition, subjects performed the SJFT at the same time of day (08:00 h). All experimental sessions were scheduled during a period with no official competitions.−2) was then calculated. Following this, duplicate measurements were taken with participants standing and wearing only briefs, as recommended by the guidelines. The average of these two measurements was used for the final analysis. Moreover, the heart rate was monitored using Automatic Blood Pressure . Before the morning test sessions, only one glass (15 to 20 cl) of water was allowed to avoid postprandial thermogenesis effects. Throughout the experimental period, participants were required to maintain their habitual physical activity and to avoid strenuous physical efforts 24 h before each test session.At the beginning of each session, the body weight, % fat, fat mass, fat-free mass, and body water were recorded using bioelectrical impedance scale to the nearest 0.1 kg  calibrated in accordance with the manufacturer's guidelines by one trained technician. The body mass index . Each subject received recommendations needed to properly complete the food diaries. Thus, they detailed the food and beverages consumed during the 3 days prior to sampling. We asked subjects to maintain their normal diet during the study period (control period). Values for nutrient intakes were obtained from a food diary for a holding period of weight and dietary restriction after 7 days. The plan was to carry out dietary restriction (−6 MJ/day) according to the report from Giannini Artioli et al. . All par Index = final HR (bpm) + HR 1 min after the end of the test (bpm)/total number of throws, where HR is the heart rate measured immediately after the test and HR 1 is the heart rate measured 1 min after the test.To perform the SJFT, three athletes of similar body mass are needed: a participant (TORI) is evaluated, and 2 other individuals receive throws (UKES). The TORI begins the test in a position between the 2 UKES who are standing 6 m away from one another. On a signal, the TORI runs to one of the UKES and employs a throwing technique called ippon-seoi-nage. The TORI then runs to the other UKE and completes another throw using the same technique. The TORI must complete as many throws as possible within the test time. The SJFT consists of three periods of 15 s (A), 30 s (B), and 30 s (C) separated by 10 s recovery intervals. Performance was determined by the total number of throws completed during each of the SJFTs . In addiBlood samples were collected using an indwelling venous catheter before (P1) (after 15 min of rest), immediately (P2), and 60 min after the exercise (P3). Then, plasma tubes were centrifuged at 3000 rpm for 20 min at room temperature and stored at −80°C until the measurement of hormonal and immune parameters . All parGH concentrations were evaluated using the ST AIA-PACK (Tosoh Bioscience). The ST AIA-PACK HGH is a two-site immunoenzymometric assay, which is performed entirely in the AIA-PACK. The intra- and interassay coefficients of variation (CV) were 0.9% and 3.6%, respectively. The limit detection was 0.07 ng/mL.−1 for testosterone and 2 ng·mL−1 for cortisol.Testosterone and cortisol concentrations were analysed by ELFA . The intra- and interassay CV were 18.2% and 5.6% for testosterone and 2.7% and 5.8% for cortisol, respectively. The limit detections were 0.1 ng·mLα were analyzed by ELISA (Enzyme-Linked Immunoassay) using commercial kits: BE53061, Hamburg, Germany, and AbC223/3, Paris, respectively. The intra- and interassay CV were 3.4% and 5.2% for IL-6 and 6.9% and 7.4% for TNF-α, respectively. All assays were carried out as advised by the manufacturer's directions. To eliminate interassay variance, all samples for each subject were assayed in the same assay.IL-6 and TNF-W-test, parametric tests were performed. Hormonal and inflammatory mediators data were analyzed using a two-way ANOVA with repeated measures (2 [diet condition] × 3 [points of measurement]). The Bonferroni post hoc test was performed whenever significant effects or a significant interaction was found using ANOVA. Energy intake, anthropometric parameters, performance of SJFT, and heart rate were tested by a paired Student's t-test. To assess the data practical significance, effect sizes were calculated as partial eta-squared, ηP2. Test-retest reliability was assessed by intraclass correlation coefficients (ICCs). The level of statistical significance was set at P < 0.05.Statistical tests were processed using STATISTICA Software . Data were reported as mean ± SD. Once the assumption of normality was confirmed using the Shapiro-Wilk P < 0.05), fat mass (P < 0.05), fat-free mass (P < 0.05), and body water (P < 0.001) with a significant decrease during CR in comparison to baseline. Likewise, macronutrient intake showed significant decrease during CR in comparison with baseline (P < 0.05) statistical differences (P < 0.05) was observed. The SJFT showed a high reliability between test-retest sessions (ICC higher than 0.87). However, heart rate  and 1 min after test (P < 0.05)) and SJFT index (P < 0.05) were significantly higher during CR in comparison to baseline  = 5.56; ηP2 = 0.54; P < 0.05), GH  = 7.33; ηP2 = 0.67; P < 0.05), and cortisol  = 5.20; ηP2 = 0.59; P < 0.05) immediately after the SJFT. Likewise, a significant diet condition effect in testosterone  = 63.35; ηP2 = 0.77; P < 0.05), GH  = 22.72; ηP2 = 0.65; P < 0.05), and cortisol concentrations  = 21.86; ηP2 = 0.77; P < 0.05) was observed (P > 0.05). Moreover, concentrations of testosterone, GH, and cortisol are significantly higher at P2 in comparison with P1 and P3 during baseline and CR (P < 0.01). CR leads to a significant decrease in concentrations of testosterone and significant increase in cortisol and GH concentrations at P1, P2, and P3 (P < 0.05).Exercise was associated with an increase in testosterone =120.3; ηP2 = 0.81; P < 0.001) and TNF-α  = 13.53; ηP2 = 0.67; P < 0.01) during P2. A significant diet condition effect on the circulating IL-6 =42.38; ηP2=0.74; P < 0.001) and TNF-α  = 32.04; ηP2 = 0.69; P < 0.001) was observed. However, the interaction diet condition × points of measurement for plasma concentrations of IL-6 and TNF-α was not significant (α is significantly higher at P2 in comparison with P1 and P3 during CR in comparison with baseline (P < 0.05). Leukocytes  = 27.53; ηP2 = 0.65; P < 0.001) and lymphocytes  = 32.43; ηP2 = 0.73; P < 0.001) during the CR were significantly lower in comparison with baseline whereas neutrophils  = 34.16; ηP2 = 0.71; P < 0.001) were significantly higher during the baseline than CR conditions  = 8.6; ηP2 = 0.67; P < 0.01) and neutrophils  = 9.7; ηP2 = 0.66; P < 0.01) increased significantly at P2 compared to P1 and remained significantly higher compared to their baseline value. However, leucocytes  = 7.77; ηP2 = 0.63; P < 0.01) were significantly lower during CR than baseline at P2.The effects of CR on inflammatory mediators during the exercise are summarized in nificant . Indeed,nditions . Lymphocα, GH, testosterone, and cortisol were higher after exercise and remained elevated 60 min after exercise during CR compared to the baseline. Likewise, leukocyte, lymphocyte, and neutrophil counts had a significant variation through exercise during the CR and the baseline conditions.The results of this study showed that 7 days of CR leads to alterations in immunological responses as well as steroid hormone during a high-intensity specific exercise in judo athletes. Indeed, IL-6, TNF-Food records, as used in this study, are considered the standard for dietary assessment and provide a quantitative account of an individual's diet during a specific period. Therefore, it is important to view the reported data of energy intake for these athletes.In this study, the caloric intake during the CR period leads to a significant decrease in body weight 4.2 ± 0.5%), which was in accordance with other athletes in combat sports , 27. The ± 0.5%, One can, then, put forward the hypothesis that the main part of the body weight loss observed during the dietary restriction is due to a body water loss. As a result, only 2% of fluid loss induces an increase in the heart rate and lowers the stroke volume, which impairs cardiac output .Our results showed an increase in plasma concentrations of cortisol, testosterone, and GH immediately after the exercise during the two experimental conditions . It has, also, been shown that weight reduction may lead to alterations in lipid profile, as it is the case in our study, which may be the consequence of the increased hormonal responses to the exercise. Our data are in agreement with previous studies that showed an increase in testosterone and cortisol immediately after exercise , 31. HowMore to the point, restrained eating may be a stressor which induced hormonal changes . These data are in accordance with the results of Anderson et al. . The lowMoreover, Loucks and Thuma  showed iα) during 7 days of CR. Judo is characterised by high-intensity, short-duration exercise and it requires the athlete to use a combination of aerobic and anaerobic capacities.To our knowledge, this study is the first to report that high-intensity exercise induced changes on proinflammatory cytokines  after CR. This report confirms cytokines as effective markers of normal recovery after intense exercise.Our results showed that plasma concentrations of IL-6 and TNF-α [α have been shown to increase by 2-3-fold after exercise [Consistently, previous reports showed that high-intensity exercise was associated with increase of proinflammatory cytokines, for example, IL-6 and TNF-α –42, whicα . Furtherα , 41. Addexercise . The majexercise .α following the 7 days of CR. These results are in agreement with previous reports that showed an alteration of the various components of the immune system. Imai et al. [Another explanation suggested that glycogen depletion, a major stimulus for IL-6 release , caused i et al.  showed ti et al. . ConsequThis study has some limitations. There was a smaller sample size of the subjects that limited our power to perform analyses. In addition, no control subjects without weight loss and without psychological stress related to an upcoming competition participated in this study. Additional studies must use proper number of weight loss and control subjects who are expending energy similar to that used for judo  but who are not participating in the competition, who are not pursuing weight loss such as open-category class athletes , and who perform either fluid restriction program or dietary energy restriction program. In addition, we did not measure direct dehydration status such as urine specific gravity. Future study needs to examine the influences of weight loss on immune parameters and somatotropic axis  in parallel with hydration status.α). Despite conducting weight loss activities to achieve an advantage in competition, the consequent weight loss might impair immune functions as well as endocrine functions, thereby depressing the overall physical condition. Therefore, it is necessary for athletes to develop safer and more effective weight loss programs. As a result, athletes, coaches, and medical staff should be sensitive to periods of increased risk of disease  and pay particular attention to recovery, the nutritional strategies, and optimal fluid intake and should monitor athletes' body weight, hydrated status, and immunocompetence to prevent their physical condition from becoming worse.In conclusion, we conclude that 7 days of weight loss programs undertaken in preparation for competition might degrade immune functions and conduct to the appearance of infectious diseases such as URTI symptoms in male judo athletes by the increase of proinflammatory cytokines (i.e., IL-6; TNF-


Usnea aurantiacoatra (Jacq.) Bory, from Fildes Peninsula, King George Island, Southwest Antarctic, were determined by radiocarbon (14C), and it is 1993–1996 at bottom and 2006–2007 at top of the lichen branch. The growth rates of U. aurantiacoatra calculated are 4.3 to 5.5 mm year−1 based on its length and ages. The comparisons show that the growth rates of U. aurantiacoatra are higher than those of U. antarctica (0.4 to 1.1 mm year−1). The growth rates of fruticose lichens are always higher, usually >2 mm year−1, than those of crustose ones, usually <1 mm year−1, in polar areas. A warming trend on Fildes Peninsula is recorded in the period from 1969 to 2010 obviously: the mean annual temperature rose from −2.75 to −1.9°C and the average temperature of summer months from 0.95 to 1.4°C, as well as the average temperature of winter months from −6.75 to −5.5°C. The alteration of lichen growth rates in polar areas may respond to the climatic and environmental changes, and the lichens may act as bio-monitor of natural condition.The ages of a fruticose lichen of  Lichens, as a unique organism with dual nature, are composed by fungi (mycobionts) and algae or cyanobacteria (photobionts) in a symbiotic relationship and became the most prominent component in vegetation The Antarctic is defined geographically as the lands and adjoining ice shelves in the south of latitude 60°S. King George Island , as the largest island of South Shetland Islands in Antarctic, locates about 120 km from the Antarctic continent and about 1100 km from the southern peak of South American continent Usnea, U. aurantiacoatra (Jacq.) Bory and U. antarctica Du Rietz, grow on Fildes Peninsula, as the most dominant members of lichens there U. aurantiacoatra collected from Fildes Peninsula were dated by 14C and hence the growth rates of this lichen are estimated to be 4.3 to 5.5 mm year−1 and are compared with those of other fruticose lichens, including U. antarctica, and also of crustose lichens living mainly in polar areas. The climatic conditions for the growths of U. aurantiacoatra and U. antarctica are considered in this work.Two species of the fruticose lichen U. aurantiacoatra studied here were collected from Fildes Peninsula, King George Island, Shetland, the southwest Antarctic by the last author C. S. Li during the 23rd Chinese National Antarctica Research Expedition in January 2007 and the specimen collection was permitted by the Chinese Arctic and Antarctic Administration, the State Oceanic Administration, People’s Republic of China. The coordinate of the collecting site is 62°12′42″ S and 58°58′55″ W, with its altitude of 43 m  and the basal part (Sample B) of a branch of r dating  and 3. T14C in the ETH Zurich. The sample was pre-treated by the ABA method (Acid-Base-Acid), i.e. kept in 4% HCl at 60°, 0.4% NaOH at room temperature and 1N HCl at 60° for one hour each step. The insoluble fraction was combusted in an Elemental Analyzer and converted to graphite using Fe as catalyst. 14C/12C ratios  were determined in the MICADAS spectrometer The sample was dated by AMS-U. aurantiacoatra had F14C >1, i.e. it had taken up anthropogenically created 14C from the atmospheric bomb testing after 1955, culminating in a doubling of atmospheric 14C in 1962/3.The sample of U. aurantiacoatra selected for dating is 60 mm in length between Sample A  to 14 (1993 to 2007) years.The branch of Sample A  and SampSample A . Sample −1.The ages of this branch are detected to be 11 to 14 years old, and the aged branch is 60 mm in length, the growth rates of the lichen are therefore 4.3 to 5.5 mm yearAs a pioneer organism, lichens can promote the weathering of rocks and pedogenesis, and create appropriate conditions for the growth of subsequent plants in the harsh environments, like desertification land and polar areas. The weathering action can be a series of physical, chemical or combined processes Deschampsia antarctica and Colobanthus quitensisThe vegetation on King George Island is the poorly developed tundra and consists almost exclusively of cryptogams, lichens and mosses, with two spices of flower plants of Neuropogon group in the genus Usnea thrive in Polar Regions and higher altitudes of temperate and tropical regions, and are often the dominant fruticose lichens in such harsh environments th and 11th Chinese National Antarctica Research Expeditions, Chen U. antarctica Du Rietz and U. aurantiacoatra (Jacq.) Bory, on Fildes Peninsula, and demonstrated that U. aurantiacoatra dominates in the local lichen vegetation.The species of U. antarctica was estimated based on the investigation of the lichens on Ardley Island close to Fildes Peninsula U. antarctica grew on the wood block of tripods there, which was built for mapping by the staff of the station of former Soviet Union in 1970, and measured to be 7 to 20 mm in length in 1993 U. antarctica produced their length of 7 to 20 mm there in 18 years (from 1975 to 1993), corresponding to a growth rate of 0.4 to 1.1 mm year−1The growth rate of U. antarctica was provided on a moraine , which is close to the Spanish Antarctic Base Juan Carlos I in South Bay, Livingston Island, South Shetland Islands, and 1 km from the coast. Ten boulders with different sizes were selected in 1991 and were marked in two designed experimental zones. Growing upright on the boulders, the thalli of U. antarctica were measured in 1991 and re-measured in January 2002. The lichens grew for 22 mm in length in 11 years , the rates of U. aurantiacoatra are much higher (4.3 to 5.5 mm year−1). The cause of the different rates of these two species of Usnea may be intrinsic. In Fildes Peninsula, U. antarctica appears at regions adjacent to the coast. When occasionally growing together with U. aurantiacoatra, it often occurs at the margin of the population of the latter U. antactica is less competitive than U. aurantiacoatra.Comparing with the growth rates of Cladonia rangiferina (L.) Weber ex F.H. Wigg. varied in different places from the North to South Hemispheres: its rates are 5.0 to 5.6 mm year−1 in spruce forest on Seward Peninsula, Alaska −1 at the site from Alakitka situated in Kuusamo, northeastern Finland −1 in tundra communities on Chukotsk Peninsula, the western Siberia −1 on the sub-Antarctic island of South Georgia The linear growth rates of C. alpestris (L.) Rabenh. were different in two localities: they are 4.3 to 5.8 mm year−1 on the Seward Peninsula, Alaska −1 on Chukotsk Peninsula, the western Siberia C. sylvatica living on the Seward Peninsula, Alaska are 5.2 to 5.5 mm year−1The growth rates of C. mitis Sandst. varied from 3.0 to 3.5 mm year−1 at Alakitka situated in Kuusamo, northeastern Finland −1 at Pine Barrens near Mauston, Juneau County, Wisconsin, USA The growth rates of U. aurantiacoatra, 4.3 to 5.5 mm year−1, are more or less similar to those of C. alpestris (3.3–5.8 mm year−1), C. rangiferina (2.7–5.6 mm year−1), C. sylvatica (5.2–5.6 mm year−1) and C. mitis (3.0–5.2 mm year−1 ), but higher than those of C. coccifera (1.6 to 2.0 mm year−1) and Evernia prunastri (L.) Ach. (2.0 mm year−1). Comparing with Bryoria spp. (rate: 13.8 mm year−1) and Ramalina reticulata (Nohden) Krempelh ( = R. menziesii Taylor) (rate: 30 mm year−1), U. aurantiacoatra grows more slowly.Considering the collected data of fruticose lichens , the rat−1, for example, the lichen of Rhizocarpon geographicum (L.) DC. living in alpine area has a growth rate below 0.5 mm year−1Aspicilia alphoplaca (Wahlenb.) Poelt & Leuckert and Trapelia coarctata (Turner ex Sm.) M. Choisy, may expand 1 mm year−1The growths of crustose lichens were investigated mainly in the temperate and Arctic areas for dating attached substrata Acarospora macrocyclos Vain. are 1.0–1.2 mm year−1 (>1 mm year−1) in Signy and Livingston Islands, Antarctic, and it is the high rate in the crustose (Placodioid) lichens in polar areas The rates of −1. For example, the rates of Rhizocarpon geographicum are 0.025–0.5 mm year−1 in Signy and Livingston Islands, and 0.1–1.4 mm year−1 in Antarctic and Arctic Caloplaca sublobulata (Nyl.) Zahlbr. are 0.82–0.95 mm year−1 in Livingston Island Buellia frigida Darb. are 0.01–0.07 mm year−1 in Dry Valleys, Cape Hallett Buellia latemarginata Darb. are 0.5–0.87 mm year−1 in Signy and Livingston Island Bellemerea sp. are 0.75 mm year−1 in Livingston Island The growth rates of most known crustose lichens in polar areas are usually less than 1 mm year−1; −1; U. antarctica, with 0.4 to 1.1 mm year−1, on Ardley Island are close to those of crustose lichens.The growth rates of fruticose lichens are always higher  and grow as upright or pendulous branches in three-dimensions with its increasing linear length, while the latter creep on the surface and expand in two-dimensions with its increasing diameter. Perhaps, the fruticose lichens may obtain more light in 3-D growth forms for their photosynthesis than the crustose lichens in a 2-D pattern.U. aurantiacoatra and U. antarctica.The lichen growths are influenced by the environmental factors, including light, temperature, precipitation, attached substance, and the interaction with other organisms at immediate vicinity. In this part, we consider the temperature data of Fildes Peninsula in relation to the growths of lichens of −1), and the 22 Southern Ocean Stations have recorded a mean rate of 0.7–1.0°C per century (0.007–0.010°C year−1). Antarctic Peninsula stations show a consistent regional rate of warming that is more than twice than the average of other Antarctic Stations The Little Ice Age is about from 1300 to 1870 We collected meteorological data available from Station Bellingshausen, Russia  The mean annual temperature (MAT) is −2.3°C at SBR from 1969 to 2010 and −2.1°C at GWSC from 1985 to 2012. The average summer monthly temperature (ASMT in January and February) is 1.6°C at SBR from 1969 to 2010 and 1.8°C at GWSC from 1985 to 2010. The average winter monthly temperature (AWMT in July and August) is −6.4°C at SBR from 1969 to 2010 and −5.9°C at GWSC from 1985 to 2010 , 5 and 6−1) in 41 years (The MATs of two stations have been rising from −2.75 to −1.9°C in the period from 1969 to 2010 (41 years .−1 (0.02–0.05°C year−1 range) is predicted to be 0.5–0.7 times larger in maritime Antarctica −1 on Fildes Peninsula (62°S) in 41 years (in this work), less than 0.056°C year−1 at Faraday Station (65°S) in 45 years higher in winter than in summer (0.030 vs 0.011year−1) on Fildes Peninsula in the last 41 years.An increase of the mean surface temperature of 1–5°C is a key prediction of climate change in the Antarctic, but at a lower rate than in the Arctic, and precipitation is expected to increase by up to 30% across the continental Antarctica Buellia frigida became fast with the increase of growth rate from 0.01 to 0.07 mm year−1, when the mean temperatures rose from −4.8 to −1.4°C in summer and −30.5 to −26.4°C in winter, with the precipitation increase from 50 to 225 mm in rainfall equivalent. The same things happened with Buellia latemarginata, whose growth rates increased from 0.5 to 0.87 mm year−1 responding to the precipitation increases from 400 to 800 mm, with the stable summer temperature (1.3 to 1.3°C) and the rising winter temperature from −9.0 to −7.0°C The lichens grow in Antarctic mainly controlled by two factors, temperature and precipitation. When the two factors changed, the lichen growth would be influenced significantly. For example, the radial growth of U. aurantiacoatra and U. antarctica (4.3 to 5.5 vs. 0.4 to 1.1 mm year−1) on Fildes Peninsula may mainly attribute to their different biological features, as mentioned before. The different attached substances of these lichens, rock vs. the wood block, may also played an important role in their growth rate. The temperature changes during the years may also influence the growth of the two species.The different growth rates of −1) from 1986 to 1999, especially cooling of 1.2°C per decade (0.12°C year−1) in summer.When we consider the warming period over the 19th century in Antarctic, on another hand, it has been slightly cooled from 1979 to 1998 U. antarctica (from 0.4 to 1.1 mm year−1) were obtained according to the assumed growth period between 1975 and 1993 U. aurantiacoatra analyzed here with its rates of 4.3 to 5.5 mm year−1 was in the period between 1993 and January 2007, with a 5-year (1993 to 1998) duration falling into the cooling interval, shorter than that of U. antarctica. This fact may also influence the different growth rates of these two lichens in some extents.The growth rates of 14C dating, a branch of Usnea aurantiacoatra (Jacq.) Bory was detected to be 1993–1996 at basal part and 2006–2007 at tip in ages. The latter estimated age of 2007 coincides with the collection year (2007) of this specimen in January of 2007, when the lichen stopped its growth. So, the dating result is accurate and reliable.By U. aurantiacoatra, found attached to the rocks in situ on Fildes Peninsula, were calculated to be 4.3 to 5.5 mm year−1. The rates of U. antarctica Du Rietz, grew on introduced substrata (wood block) on Ardley Island, were estimated to be 0.4 to 1.1 mm year−1 previously.Based on the length (60 mm) and ages (11 to 14 years) of this branch, the growth rates of −1, than those of crustose lichens, usually <1 mm year−1, in polar areas.The growth rates of fruticose lichens are always higher, usually >2 mm year−1).The mean annual temperature have been rising from −2.75 to −1.9°C in the period on Fildes Peninsula from 1969 to 2010, and the average summer monthly temperature (December–February) rising from 0.95 to 1.4°C, as well as the average winter monthly temperature (June–August) rising from −6.75 to −5.5°C. A warming trend on Fildes Peninsula is obvious, especially with the temperatures rising higher in winter than in summer (0.030 vs 0.011°C yearThe climate and environment changes influence the lichen growths in polar areas significantly. When the ages of lichens are detected exactly, the changes of lichen growth in these ages may respond to the climate and environment changes, and the lichens may play a bio-monitor of natural condition therefore.


AbstractDraconariusjiangyongensis  is described for the first time from Xinning County, Hunan Province, China. Morphological descriptions and illustrations of both sexes of this species are given in this study. The placement of this species in Draconarius is doubted.The male of  Draconarius Ovtchinnikov, 1999 is distributed in Central and East Asia and has a high level of species diversity with 244 species described to date  was first described as a member of the genus Coelotes Blackwall, 1840 based on six female specimens from Jiangyong County, Hunan Province, China .Specimens were examined with an Olympus SZX16 stereomicroscope and an Olympus BX53 compound microscope. Photos were taken with a Canon PowerShot G12 digital camera mounted on an Olympus BX53 compound microscope. Both the male palp and the female epigyne were examined and illustrated after being dissected from the spider bodies. All specimens examined in this study are deposited in the College of Life Sciences, All measurements are given in millimeters. Eye diameters are taken at the widest point. Leg measurements are given as: total length . Abbreviations used in the text are as follows: anterior median eyes AME anterior lateral eyes ALE median ocular area MOA posterior median eyes PME posterior lateral eyes PLETaxon classificationAnimaliaAraneaeAgelenidaeCoelotesjiangyongensisDraconariusjiangyongensisCoelotes);  : HNU), Jiangyong County, Hunan Province, China, 1 October 1991, Liansu Gong leg.; paratypes, 4♀ (HNU), same data as holotype.Holotype, 1♀ , Tianyixiang , 21.11.2014; 3♀, 2♂ (HNU), same locality as above , 22.11.2014; 1♀, 1♂ (HNU), Cave Feiliandong , 23.11.2014; 1♀ (HNU), Bajiaozhai , 24.11.2014; 5♀, 8♂ (HNU), Peak Lajiaofeng , 27.11.2014. All specimens are collected by hand picking by Haiqiang Yin, Cheng Wang, Bing Zhou, Jiahui Gan and Yuhui Gong.Draconariusjiangyongensis can be distinguished from other Draconarius by the presence of a vase-shaped septum of epigyne  in having a simple conductor, an embolus arising at approximately 10 o’clock  and the short cymbial furrow   Figs , 3B, D.Male. Total length 8.7. Carapace 4.1 long, 3.1 wide; opisthosoma 4.5 long, 2.9 wide. Clypeus height 0.15. Cephalic part much elevated from the thoPageBreakracic region. Cervical and radial grooves greyish-black , III 14.0 , IV 17.8 . Opisthosoma with dorsal pattern composed of several chevrons patterns . Leg measurements: I 14.3 , II 12.2 , III 10.8 , IV 14.1 . Promarginal and retromarginal teeth of chelicera and the dorsal pattern of opisthosoma are the same as male  and short copulatory ducts in the female; the large patellar apophysis, the short and prolaterally originating embolus and the short cymbial furrow (less than 1/3 length of the cymbium) in the male. It differs from Coelotesatropos  by the presence of septum. It also differs from the type species and many other species of Draconarius , Draconariuslatellai Marusik & Ballarin, 2011 and so on) by the number of cheliceral teeth The China (Hunan).


Cow’s milk (CM) oral immunotherapy has proven to be effective as a treatment for persistent CM allergy. Allergic reactions to goat and sheep cheeses have been described in almost 25% of patients desensitised to CM, indicates that immunotherapy is specific for each milk.To describe changes of goat’s IgE and sheep’s IgE in patients who tolerated CM after a CM oral immunotherapy.24 children aged between 4 and 13 who achieved tolerance to milk after CM immunotherapy were included. Specific IgE to cow’s caseine, sIgE to whole goat’s milk and sIgE to whole sheep’s milk at baseline and at 24 months follow up were analised (CAP-Phadia). The patientes were classified in two groups: high risk (17 patients) and low risk (7 patients) based on the premedication need, number of adverse reactions and time to achieve a complete tolerance to CM.The median of Specific IgE to CM casein, goat’s whole milk, sheep’s whole milk a baseline were 24.3KU/L, 31.2KU/L and 27.4KU/L and 9.5KU/L, 16.9KU/L and 17.3KU/L after 24 months of follow up .In the high risk patients the median of Specific IgE to CM casein, goat’s whole milk, sheep’s whole milk were 33.7KU/L, 43.7KU/L and 37.1KU/L at baseline and 13.1KU/L, 23.9KU/L and 23.6KU/L after 24 of follow up . In contrast, in the low risk group the specific IgE to CM casein, goat’s whole milk, sheep’s whole milk did not shown a significant decrease after 24 of follow up. 4 patients showed a different patron referred to IgE-goat’s milk and IgE-sheep’s milk compared with CM, independently of risk group.We found a decrease of IgE levels to whole goat´s and sheep´s milk after oral immunotherapy to CM.


Our method combining with the dynamical diffraction theory simulation can uniquely provide unit cell dimensions layer by layer, and is applicable to thicker samples.Interfacial strains are important factors affecting the structural and physical properties of crystalline multilayers and heterojunctions, and the performance of the devices made of multilayers used, for example, in nanowires, optoelectronic components, and many other applications. Currently existing strain measurement methods, such as grazing incidence X-ray diffraction (GIXD), cross-section transmission electron microscope, TEM, and coherent diffractive imaging, CDI, are limited by either the nanometer spatial resolution, penetration depth, or a destructive nature. Here we report a new non-destructive method of direct mapping the interfacial strain of [001] Si In recigg’s law , three r0.7Ge0.3/Si, is prepared by using a germanium silicide target to grow a thin-film of Si0.7Ge0.3 on silicon substrate by molecular beam epitaxy, MBE, in National Nano Device Laboratory, NDL. In addition, before the epitaxial process, the Si wafer clearance is a standard manufacturing procedure, i.e., the surface particles and native SiO2 are removed. An energy dispersive spectrum, EDS, is employed to measure compositions versus depth  of 0.7Ge0.3 thickness, dSiGe (~57 nm). The c-axes of the Si0.7Ge0.3 and silicon substrate are about 5.5344 and 5.4345 Å, because of two distinct 004 peaks  and interface (σinterface) are about 0.9 and 1.1 nm, respectively. Moreover, through the image of high-resolution transmission electron microscope, HRTEM, the local probe of the Si0.7Ge0.3 thickness is about 59.9 nm , National Synchrotron Radiation Research Center (NSRRC). For the BSD, 004/202, the sample is mounted at the center of an 8-circle diffractometer. For 12 keV X-rays, after the alignment for the symmetric Bragg reflection, 004, and the surface reflection, 202, the intensity distributions of vertical (tth) and horizontal (beta) scans of the surface 202 reflection are measured with the slit sizes 10 mm(H) × 0.1 mm(V) and 0.1 mm(H) × 0.1 mm(V), respectively.The BSD diffraction experiments of Si0.7Ge0.3 and Si respectively. The other two peaks, p2, p3 and the oscillating background are the Kiessig fringes31112The tth-scan of surface diffraction, 00.7Ge0.3 thin-film and silicon substrate so that the simulated intensity distributions fit the measured ones. Namely, the elements of the metric tensor, σ, defined below will be determined,The simulations are started from finding unique solutions for the three lattice unit vectors, x, ay, az), x, by, bz) and x, cy, cz). The peak positions of the tth- and beta- scans of the three surface reflections, 202, 0σ.where 0.7Ge0.3, denoted as σ1. Similarly, the measured tth and beta of p4 from σ2. For clarity, the tensor elements of σ are labeled as  and σ2 (Si) at corresponding depth, x, using the Boltzmann function as an approximation:As the samples are high-quality crystals with narrow peak widths , the dyn31112EDS data  and VegaEDS data . Namely,0 is the thickness of the Si0.7Ge0.3, determined from the Bragg peak at p1. The two constants, lat1 and lat1, are the corresponding elements of the tensors, σ1 and σ2, respectively. For example, for Β(x) at depth, x, c0 is the changing rate of lattice parameters in depth, which is determined by fitting the simulations to measured intensity distributions.where the symbol, x, is the depth from the crystal surface, and xc0 of the lattice parameters for the Si0.7Ge0.3/Si sample of a thousand atomic layers along [001], the multilayer dynamical theory of X-ray diffractionnth atomic layer are calculated as l = 1) and bottom boundary (l = 2) of the nth layer considered. For multilayers, the coefficients, nth-to-(n + 1)th layers, and the bottom (exit) surface of the sample. This leads to the diffracted E-fields given in To determine the changing rate Im, as below:Moreover, due to the shallow diffraction angles of the surface reflections with respect to the crystal surface, part of the diffracted beams are absorbed by the crystal. Therefore a fitting function nth layer’s top boundary for the mth reflection. Note that m = 0 is for the forward diffracted beam O, m = 1 for the symmetric Bragg reflection G, and m = 2 for the surface diffraction L, and nth layer. nth layer’s top boundary for the mth reflection. The terms, Δθ (~0.007 deg.) and Δϕ (~0.02 deg.) are the vertical and horizontal divergences, respectively on the beamline BL17B1. The symbol, M is the total number of atomic layers of the simulation i.e, M = 1000. The position vector, where c0 = 0.6 for these cases. When c0 < 0.6, the measured intensity distributions of the interference between the diffraction from the Si0.7Ge0.3 and from the Si substrate cannot be matched by the simulated curves. When c0 > 0.6, the simulated peak intensities at p1 and p4 will be off the measured values. With c0 = 0.6, the nine elements of the σ tensor as functions of depth in Å are determined, which are shown in The best fit between the simulated intensity distributions to the measured ones is reached when 0.7Ge0.3, are grown on the silicon wafer, the a- and b-axis of the thin-film, Si0.7Ge0.3, are bound to the silicon substrate. To relax the strain, the c-axis of the thin-film, Si0.7Ge0.3, is longer than a- and b-axis, and the structure became rectangular rather than cubicFurthermore, the six lattice parameters can be also calculated from the σ-tensor. The results are shown in 0.7Ge0.3/Si are successfully obtained. This method can be used for strain mapping of a wide variety of multilayer crystal systems .


Objective. The study aimed to evaluate the influences of coadministration of antiepileptic drugs (AEDs) on an antimalarial candidate 99/411 pharmacokinetic (PK) profile. Method. For this, single oral dose PK drug interaction studies were conducted between 99/411 and FDA approved AEDs, namely, Phenytoin (PHT), Carbamazepine (CBZ), and Gabapentin (GB) in both male and female SD rats, to assess the coadministered and intersexual influences on 99/411 PK profile. Results. Studies revealed that there were no significant alterations in the PK profile of 99/411 upon PHT and CBZ coadministration in both male and female rats, while systemic exposure of 99/411 was significantly increased by about 80% in female rats upon GB coadministration. In terms of AUC, there was an increase from 2471 ± 586 to 4560 ± 1396 ng·h/mL. Overall, it was concluded that simultaneous administration of AEDs with 99/411 excludes the requirements for dose adjustment, additional therapeutic monitoring, contraindication to concomitant use, and/or other measures to mitigate risk, except for GB coadministration in females. These findings are further helpful to predict such interactions in humans, when potentially applied through proper allometric scaling to extrapolate the data. In a number of clinical situations, concomitant use of two or more drugs is imperative. In particular in chronic disease treatment course, patients must take more than one drug for an extended period of time. In this course, many drug interactions have received a great deal of attention to avoid any side effects to the patients. The greater the number of drugs prescribed to a patient, the greater the risk of drug interactions. In an epidemiological study, conducted at a community hospital to investigate the clinical relevance of drug-drug interactions (DDIs), it was reported that incidence of interactions raises up to 7% in those patients taking 6–10 drugs and 40% in those taking 16–20 drugs a day .Many of the major pharmacokinetic (PK) interactions between drugs are due to cytochrome 450 (CYP450) enzymes, whose genetic expressions are being affected (induction or inhibition) by previously or simultaneously administered drugs. In addition, such interactions may exist at the level of absorption, distribution, clearance, and transporters. Such interactions are investigated in terms of the PK parameter changes of the concerned drug. These substantial changes in PK parameters may be used to indicate clinical importance of DDIs, when substrate and interacting drugs are likely to be given simultaneously either for short periods or chronically for an extended periods of time during the combinational therapy. in vivo drug interaction studies are usually carried in experimental animals because of the difficulties to conduct these studies in humans at this stage [The present study aimed to evaluate the influences of coadministration of antiepileptic drugs (AEDs) on an antimalarial candidate 99/411 pharmacokinetic (PK) profile. 99/411 is a novel 1, 2, 4 trioxane derivative, antimalarial drug candidate with peroxide scaffold essential for its pharmacological activity . This mois stage , 7. Thesis stage , provideIn this study, our investigational drug 99/411 was considered as a substrate and the interacting agents chosen were FDA approved antiepileptic drugs (AEDs) including Phenytoin (PHT), Carbamazepine (CBZ), and Gabapentin (GB). The rationale behind selecting AEDs as interacting drugs is being strengthened by the fact that the seizures (characteristics of epilepsy) are one among the common feature of malaria regardless of its severity , 11. Thu Sprague-Dawley (SD) rats provided by National Laboratory Animal Centre (NLAC), CDRI. Pure (>99%) reference standards of 99/411 and 97/63 (used as IS) Figures  were obtHealthy male and female SD rats obtained from NLAC of CDRI, India, were weighed 200–250 g on the day of dosing. Prior to the studies, animals were acclimatized for 4 days in proper ventilated polypropylene cases in standard laboratory conditions with regular 12 h light-dark cycle, temperature (22 ± 2°C), and relative humidity (55 ± 5%). Standard pellet laboratory chow and water were allowed ad lib. Guidelines approved by the “Animal Experimentation Ethics Committee” and Good laboratory practice (GLP) were followed throughout the animal experimentation.n = 3). Studies were conducted separately on male and female SD rats to assess the intersex differences in the drug interaction pattern. Subjects were given 12 mg/Kg of 99/411 (control) and 99/411 followed by coadministration of AEDs (42 mg/Kg each) in separate sets of experimental animals. Blood samples for PK analysis were collected before dosing and at the following time points 0.083, 0.33, 0.75, 1, 1.5, 2, 4, 6, 8, 12, 24, 36, 48, 60, 72, and 96 hours after dose.The studies were single dose oral PK, designed to estimate the effect of concurrent administration of AEDs on PK profile of 99/411. Individual studies were conducted for each AED coadministration with 99/411. Each study was carried out in two groups of experimental animals, every group comprising three animals  at 12 and 42 mg/Kg dose, respectively. Both the formulations were subjected to quality control (QC), stability, and homogeneity test to ensure the strength before dosing. The volume factor for both the formulations was 1 mL/Kg.µL of IS was spiked into each study samples followed by a double step liquid-liquid extraction procedure using in n-hexane to extract the drugs and IS. Plasma concentration was determined by using a fully validated LC-MS/MS method. Mass spectrometric detection was performed on an API 4000 LC-MS/MS mass spectrometer  with Analyst 1.4 software. Product ion transitions , at m/z 412.3 to 185.1 and 418.2 to 119.1 were used for quantification of 99/411 and IS respectively. The assay was linear over the range 1.56–200 ng/mL with LOQ 1.56 ng/mL [r2) were >0.99 for standard curves generated. Precision and accuracy of the method was determined by analyzing QCs at 1.50, 80 and 160 ng/mL. All accuracies were within 15% of the nominal concentrations with standard deviation of <8%.Blood samples collected for quantitative estimation of 99/411 were processed by centrifugation within 30 min of sample collection to obtain plasma and were stored at −70°C till analysis. 10 ∞0–), maximum plasma concentration (Cmax), time to attain Cmax (Tmax), elimination half-life (T1/2), mean residence time (MRT), and relative bioavailability (RB), when 99/411 was given alone and coadministered with AEDs in male and female SD rats. These PK parameters were calculated by noncompartmental analysis using Winnonlin software . The estimated PK parameters for 99/411 alone and coadministered with AEDs were statistically compared using two-tailed Student's t-test. In all the tests, a probability level of significance was kept at α = 0.05. Results were expressed as mean ± SD. RB was calculated asThe primary endpoints for these studies were area under the curve . The plasma concentration-time profiles  for 99/411 in male and female rats have been shown in To evaluate the coadministered influences of AEDs on PK profile of 99/411, its baseline PK profile was generated at 12 mg/Kg in healthy male and female rats. After oral dose administration, AUCfference . PK paraT1/2, Tmax, Cmax, AUC∞0–, and MRT for 99/411 upon coadministration of 99/411 with PHT were 2.10 ± 0.30 h, 7.00 ± 1.00 h, 489.00 ± 166.4 ng/mL, 2521.93 ± 619.89 ng·h/mL, and 17.58 ± 2.85 h, while for female rats values were 4.65 ± 0.67 h, 4.00 ± 1.18 h, 493.85 ± 73.14 ng/mL, 2160.70 ± 488.58 ng·h/mL, and 12.04 ± 1.00 h, respectively. PK profiles after oral coadministration of 99/411 with PHT have been given in In male rats, mean values of P values which were found >0.05 for respective parameter. However, RB was found to be enhanced in male rats, while a decrease was seen in female rats (n = 3) for 99/411 upon PHT coadministration in male and female rats have been shown in Intersex statistical analysis of PK parameters for 99/411 upon coadministration with PHT in male and female rats revealed that no significant difference existed between male and female PK profiles in terms of ale rats . The plaT1/2, Tmax, Cmax, AUC∞0–, and MRT for 99/411 upon coadministration of 99/411 with CBZ were 3.16 ± 0.49 h, 6.50 ± 0.50 h, 495.25 ± 58.42 ng/mL, 2534.84 ± 299.69 ng·h/mL, and 13.84 ± 1.56 h in male rats, while 3.49 ± 1.09 h, 4.00 ± 1.18 h, 484.74 ± 60.50 ng/mL, 2095.20 ± 298.73 ng·h/mL, and 12.17 ± 0.98 h for female rats, respectively  for 99/411 upon CBZ coadministration in male and female rats have been shown in Intersex statistical analysis of PK parameters for 99/411 upon coadministration with CBZ in male and female rats revealed that no significant difference existed between male and female PK profiles in terms of ale rats . The plaT1/2, Tmax, Cmax, AUC∞0–, and MRT for 99/411 upon coadministration of 99/411 with GB were 3.16 ± 0.49 h, 6.50 ± 0.50 h, 495.25 ± 58.42 ng/mL, 2534.84 ± 299.69 ng·h/mL, and 13.84 ± 1.56 h in male rats, while 5.02 ± 1.09 h, 2.00 ± 1.11 h, 449.64 ± 77.95 ng/mL, 4560.86 ± 1396.05 ng·h/mL, and 14.05 ± 1.56 h for female rats, respectively. PK profiles after oral coadministration of 99/411 with GB have been given in Mean values of P values. However, RB was found to be decreased in male rats, while an increase was seen in female rats (n = 3) for 99/411 upon GB coadministration in male and female rats have been shown in Intersex statistical analysis of PK parameters for 99/411 upon coadministration of 99/411 with GB in male and female rats revealed that no significant difference existed between male and female PK profiles in terms of ale rats . The plaThis study was conducted to evaluate the influence of concurrent administration of 99/411 with FDA approved AEDs. Keeping in view, the effect of species and sex differences on the PK profile of concerned drugs, study was conducted in healthy male and female SD rats , 13. FirP values > 0.05) from baseline PK profile, indicating that PHT coadministration has little influence on PK parameters of 99/411. RB of 99/411 was observed to be enhanced by about 20% in male rats, while a decrease of about 13% was found in female rats. When PK profiles of male and female rats were statistically compared to evaluate intersex effect, it was observed that significance difference existed for T1/2 values, while for other PK parameters no significance difference existed  of 99/411 in male rats, while an increased AUC∞0– was found for female rats. In terms of RB, RB of 99/411 was significantly increased by about 80% in female rats, while no effect was observed in male rats. Thus, a significant difference was observed intersexually in the interaction pattern influencing the RBs. The RB was almost unaltered in male rats, while significantly enhanced in female rats. In male rats, there was a shift in Tmax from 2 ± 0.00 to 6.50 ± 0.50 h, indicating that GB coadministration interacted at absorption level, which might have reduced absorption rate of 99/411; however, systemic exposure was unaltered. Conversely, in female rats, Tmax was shifted from 4.50 ± 1.53 to 2.00 ± 1.11 indicating fast absorption of 99/411 upon GB coadministration which might have resulted into enhanced systemic exposure of 99/411.Like PHT and CBZ, GB is widely used AED, which possesses quite desirable PK properties. It does not bind to plasma proteins nor is metabolized and does not induce/inhibit liver enzymes, reducing the likelihood of drug interaction with other drugs. Although this drug is rapidly absorbed in small intestine by carrier mediated transport system, which is saturable . This lesorption . Study wn = 3); results represented herein do not negate the variations in the interaction pattern. These findings could not be directly utilized to estimate and predict existence of similar interactions in humans, unless predictions are not rationalized through allometric scaling approaches, keeping in view all the anatomical, physical, and chemical variations across species.Overall, these studies explicitly indicated that coadministration of these AEDs does not exert any significant effect on PK of 99/411, except for GB coadministration in female rats. A similarity was observed in the interaction pattern of PHT and CBZ, where an increase in the RB of 99/411 was reported in male rats, while a decrease in female rats, which was nonsignificant though. In contrast, opposite pattern was found for GB coadministration, where a significant increase of about 80% was reported in female rats, while no effect was reported in male rats. However, these studies were conducted on a randomized selection basis of animal group (


Pulmonary adenocarcinoma (PA) is the most common histologic type of primary lung cancer. Generally, adenocarcinoma was composed by five major components. The present study aimed to evaluate changes in the composition of adenocarcinoma components as the tumor grows; in addition, to analyze the correlation between the occupancy rates of histologic components of the tumor in regard to prognosis.Pathologic data were retrospectively evaluated for 206 patients who underwent curative resection of PA. We investigated how histologic component occupancy rates changed as tumor size and N stage increased. To evaluate local invasiveness, the major components of the present group and absent group of pleural invasion, lymphatic invasion, and vascular invasion were compared.P = 0.006, P < 0.001) ; however, the percentage of lepidic components decreased (P < 0.001). In cases with a solid component and a micropapillary component, a gradual increase was found with an increase N stage ; however the percentage of lepidic components decreased (P < 0.001). Average differences of histologic components dependent upon whether pleural, lympathic and vascular invasion were present, the difference of micropapillary and lepidic components were statistically significant. With logistic regression analysis, as the occupancy rate of the lepidic component increased, the probability of pleural invasion, lymphatic invasion, and vascular invasion decreased; in cases with a micropapillary component, as the occupancy rate of increased, the probability of lymphatic invasion and vascular invasion increased. In multivariate analysis using the Cox propotional hazards model, the occupancy rates of acinar, micropapillary and lepidic  components were significantly associated with recurrence.The mean percentages of acinar and solid components significantly increased with an increase in size (The lower the occupancy rate of a lepidic component and the higher the occupancy rates of acinar, solid, and micropapillary components, the likelihood of tumor progression increased. In addition, as the occupancy rate of a lepidic component decreased and a micropapillary component increased, local invasiveness and recurrence rate increased; thus, increasing the probability of a poor prognosis. Pulmonary adenocarcinoma (PA) is the most common histologic type of primary lung cancer . Since NThe majority of primary adenocarcinomas that occur in the peripheral lung have the radiologic appearance of a ground glass opacity nodule (GGN) at the incipient stage; in most cases, the lesion becomes more solid as the tumor grows , 5. MoreThe most important prognostic factor of non-small cell lung cancer is its anatomical stage; this staging is commonly determined by the American Joint Committee on Cancer (AJCC) TNM stage . HoweverFrom March 2011 through September 2013 at Seoul St. Mary’s Hospital in Korea, 407 patients were diagnosed with non-small cell lung cancer and underwent pulmonary resection that entailed more than a lobectomy for an attempt to achieve a radical cure. Among these 407 patients, 275 were diagnosed with adenocarcinoma. The following patients were excluded from the evaluation: treatment with neoadjuvant chemotherapy preoperatively; diagnosed with atypical adenomatous hyperplasia (AAH), adenocarcinoma in situ (AIS), or minimally invasive adenocarcinoma (MIA); had multiple lesions, staging was not possible because a lymph node dissection was not performed; or the occupancy rates of histologic components were not recorded. Ultimately, 206 patients were included in this retrospective chart review. This study was approved by the Institutional Review Board of Seoul St. Mary's Hospital (The Catholic University of Korea).The five components of adenocarcinoma were recorded in 5% increments. The recording method was followed via the recommendations of IASLC/ATS/ERS . If the P < 0.05.The averages of the respective component occupancy rates in each tumor group classified by size and N stage were compared by ANOVA and the Kruskal Wallis H test; the averages of the respective component occupancy rates of each component classified with pleural invasion, lymphatic invasion, and vascular invasion were comparatively analyzed via the t-test. In addition, logistic regression was used for the analysis of the factors influencing pleural invasion, lymphatic invasion, and vascular invasion. Statistical significance was set at The mean patient age was 63.13 years (range: 33–85 years). A total of 91 patients (44.2%) were men, and 115 patients (55.8%) were women. In regard to the location of tumor onset, it was central in 15 cases (7.3%) and peripheral in 191 cases (92.7%); thus, the tumor onset was primarily in the peripheral area. In regard to stage, it was IA in 114 cases (55.3%), IB in 36 cases (17.5%), IIA in 22 cases (10.7%), IIB in 9 cases (4.4%), IIIA in 21 cases (10.2%), IIIB in 1 case (0.5%), and IV in 3 cases (1.5%).Classification by tumor size was in accordance with T stage criteria; the tumor size was < 2 cm in 80 cases (38.9%), ≥ 2 cm and < 3 cm in 65 cases 31.6%), ≥ 3 cm and < 5 cm in 8 cases (3.8%), and ≥ 7 cm in 2 cases (0.9%). In regard to N stage, it was N0 in 164 cases (79.6%), N1 in 22 cases (10.7%), and N2 in 20 cases (9.7%). Pleural invasion was present in 50 cases (24.3%), lymphatic invasion was present in 95 cases (46.1%), and vascular invasion was present in 40 cases (19.4%) . In cases with a solid component, it also increased as the size increased: < 2 cm, 3.1%; ≥ 2 cm and < 3 cm, 11.8%; and ≥ 3 cm, 13.1% (P < 0.001). However, in cases with a lepidic component, it decreased as the size increased: < 2 cm, 49.6%; ≥ 2 cm and < 3 cm, 30.0%; and ≥ 3 cm, 14.9%. In micropapillary component cases, it increased as the size increased: < 2 cm, 2.3%; ≥ 2 cm and < 3 cm, 4.8%; and ≥ 3 cm, 7.1%; however, the difference was not statistically significant (P = 0.143). In cases with a papillary component, there was no significant change (P = 0.138) ; ≥ 2 cm and < 3 cm, 65 31.6%), and ≥ 3 cm, 61 (29.6%). We compared the change in the average histologic component occupancy rate in accordance with the increase of size. In cases of a predominant acinar component, when the size was < 2 cm, it was 35.6%; it was 35.8% when the size was ≥ 2 cm and < 3 cm. However it increased significantly to 48.52% when the size was ≥ 3 cm . In cases with a micropapillary component, a gradual increase was also found: N0, 3.5%; N1, 6.6%; and N2, 11.0% (P < 0.001). However in cases with a lepidic component, a gradual decrease was found: N0, 39.9%; N1, 7.5%; and N2, 6.3% (P < 0.001); no significant changes were found in cases with an acinar (P = 0.051) or papillary component (P = 0.053) ; micropapillary component, 7.8% (P < 0.001); and solid component, 15.6% (P < 0.001). In cases with a lepidic component, it was 39.8% in the group without pleural invasion and 13.4% in the group with pleural invasion; thus, showing a statistically significant lowering-aspect (P < 0.001) ; and cases with a solid component, 12.2% (P < 0.001). In cases with a lepidic component, it was 44.2% in the group without lymphatic invasion and 20.2% in the group with lymphatic invasion; thus, showing a statistically significant lowering aspect (P < 0.001) , in cases with a micropapillary component, it was 8.5% (P = 0.002), and in cases with a solid component, it was 15.0% (P = 0.027). In cases with a lepidic component, it was 38.5% in the group without vascular invasion and 11.0% in the group with vascular invasion; thus, showing a statistically significant lowering-aspect (P < 0.001) ) and solid components increased ; a similar decrease was found as the occupancy rate of the lepidic increased . In cases of lymphatic invasion and vascular invasion, only the micropapillary and lepidic types showed significant results. As the percentage of the micropapillary component increased, the likelihood of lymphatic invasion  and vascular invasion ) increased. As the percentage of the lepidic component increased, the likelihood of lymphatic invasion ) and vascular invasion ) decreased . During follow up, 35 patients (17%) of all patients n = 206) and 16 patients of stage I patients (n = 150) experienced recurrence. In stage I, 3-year disease free survival was 86.6% . In cases with an acinar component, the growth rate increased as the N stage increased; however, the increase did not reach statistical significance. In summary, we found that as the tumor progressed, the occupancy rate of the lepidic component decreased while that of the solid component increased.Pleural invasion, lymphatic invasion, and vascular invasion are known prognostic factors of lung cancer. Cases with pleural invasion are considered to be one of factors influencing prognosis because pleural invasion is one of the factors included in the TNM stage . HoweverOur data were relatively recent data, so we couldn’t analyzed 5-year overall survival. However, we performed 3-year disease free survival analysis. In multivariate analysis, an increase of micropapillary component occupancy and an decrease of lepidic component occupancy were significantly associated with recurrence. This results showed that the occupancy rates of micropapillary and lepidic components are related with prognosis.All of the foregoing results contributed somewhat to the prediction of the characteristics and changes of the typical histologic components of pulmonary adenocarcinoma. Each component would have different characteristics, which could influence the clinical course. If the occupancy rates of these histologic components were analyzed in detail and the characteristics in accordance with the change in the occupancy rates were revealed, individualization of a treatment protocol for adenocarcinoma could defined; thus, helping to maximize the benefits of the treatment.Our study had several limitations. It is retrospective and single center. The patient sample was relatively small; this particularly impacted categorization by tumor size. Even though the classification was made in compliance with T stage criteria of TNM stage, the number of patients in the group > 5 cm was only 10; thus, the group was combined with the group ≥ 3 cm and < 5 cm. If the patient sample was larger, more size subdivisions could be made and more accurate results could have been attained; furthermore, this would increase the significance of the study. Long-term follow-up was not conducted. Long-term follow-up regarding the influence of each histologic component could provide a more detailed analysis of the recurrence rate and mortality.In most cases, pulmonary adenocarcinoma consists of various histologic components and the occupancy rate of each component changes as the tumor progresses. The lower the occupancy rate of a lepidic component and the higher the occupancy rates of acinar, solid, and micropapillary components, the likelihood of tumor progression increased. In addition, as the occupancy rate of a lepidic component decreased and the occupancy rate of a micropapillary component increased, local invasiveness and recurrence rate increased; thus, increasing the probability of a poor prognosis.