Planting patterns affect nitrogen (N), phosphorus (P), and potassium (K) acquisition and distribution in maize and soybean under intercropping conditions. Here we reveal that strip relay-intercropping increases the N, P, and K uptake and distribution across plant organs  of maize and soybean, accelerates the dry-matter production of intercrop-species, and compensates the slight maize yield loss by considerably increasing the soybean yield. In a two-year experiment, soybean was planted with maize in different planting patterns  of relay-intercropping, both planting patterns were compared with sole cropping of maize (SM) and soybean (SS). As compared to SI, SII increased the N, P, and K accumulation in each organ of soybean by 20, 32, and 18 (root) %, 71, 61, and 76 (straw) %, and 68, 65, and 62 (seed) %, respectively, whereas decreased the N, P, and K accumulation in each organ of maize by 1, 4, and 8 (root) %, 1, 10, and 3 (straw) %, and 5, 10, and 8 (seed) %, respectively. Overall, in SII, relay-cropped soybean accumulated 91% of total nutrient uptake (TNU) of sole soybean plants, and relay-cropped maize accumulated 94% of TNU of sole maize plants. It is important and continues to be broadly practiced not only under tropical regions2, but in temperate regions also3, resulting in higher seed yields4. The maize soybean relay-intercropping is the major planting system, especially under rainfed regions7. As compared to intercropping systems, the advantage of relay-intercropping system is higher because in intercropping systems both crops almost have the similar growth periods and they required high amount of inputs to produce higher intercrop yields, whereas under relay-intercropping system both crop species have different growth periods and have complementary resource use in time. In addition, for maize soybean relay-intercropping system (MSR), the land equivalent ratio (LER) often reaches 1.7–1.8 when both crops were planted at their optimal planting density8, which increases its popularity among farmers, especially among small farmers. In MSR, spring maize is sown as two rows narrow strips in the first or second week of April, while two rows of soybean are sown in the fellow wide rows in the mid of June. Maize is subsequently harvested in the mid of August and soybean is harvested in the first week of November9. Previously, researchers have obtained higher values of LER under MSR11. The reasons for high LER are as follows: (a) increase in the gap between maize and soybean rows by narrowing the distance between maize rows12; (b) use of similar planting density between sole and intercropping system by adjusting the plant distance  in every row under intercropping conditions, which compensates for decreased row number10; and (c), optimum light distribution in maize and soybean rows by developing compact varieties of maize which enables sun light to reach maximum at soybean canopy13. Presently this system has been used in many provinces of China, such as Ningxia, Henan, Guizhou, Shandong, Sichuan, etc.15. The maximum intercrop yields in every province were utilized for comparison to estimate LER, e.g., 9000 and 3000 kg ha−1 for maize and soybean, respectively in Shandong province. This method reached LER of 1.8 under relay-intercropping system11, which is much higher as compared to the averaged value of LER 1.22 ± 0.0216 and 1.3017. In relay-intercropping system, maize had a yield advantage due to the better growth and border rows effect, and almost produced equal or higher seed yield than sole maize yield11, whereas soybean is the later grown crop of this system, as the result of crop competition with maize, the early growth of soybean is slower than sole soybean18. Previously, it has been reported that during the co-growth period soybean suffered from heavy maize shading which negatively affected the plant morphology19, physiology13 and carbon production20. In addition, relay-intercropped soybean attained lower photosynthetic rate21 and biomass per plant than sole cropped plants22. However, soybean plants exhibited fast recovery growth after the harvest of maize crop, and the severe competition for light and nutrients from maize plants during the co-growth period is compensated after the maize harvesting because the ability of soybean recovery growth was found to be stronger in MSR23. Moreover, in later stages of growth, soybean under relay-intercropping system has the better access to land, nutrients, light, and water resources per plant in comparison with sole cropping system8. In past, it has been found that the stronger recovery growth of relay intercropped crop species related to higher nutrient uptake24 and radiation use efficiency of soybean plants under relay-intercropping system14. In addition, this type of strong competition during the co-growth period and then relaxed competition during the later growth stages recognized as ‘competition recovery production principle25.Relay-intercropping systems are used worldwide since these can increase the land use efficiency and nutrient use efficiency26. However, no experiment has been carried out to investigate the major nutrient  uptake and distribution among different plant parts of maize and soybean under different planting patterns in MSR. Some researchers have documented the nitrogen (N) and phosphorus (P) uptake in maize under wheat-maize relay intercropping system under different row configuration27, but the effects of different planting patterns on nutrient uptake and distribution in maize and soybean are not clearly understood. Therefore, it is important to investigate the nutrient uptake and distribution pattern in maize and soybean for sustainable agriculture production. In addition, relay-intercropping systems are different from the intercropping systems in co-growth periods of intercrop species, and planting pattern plays a vital role in increasing the intercrop advantage28. Up till now, the nutrient uptake and distribution pattern among different plant parts of maize and soybean under MSR remain unclear. Therefore, investigating the effects of different planting patterns on nutrient uptake and distribution in intercrop species is important in making nutrient decisions for intercrop species and to maintain agriculture sustainability in MSR. This experiment aims (i) to investigate the effects of different planting patterns on nutrient uptake and distribution in maize and soybean at plant and organ level; (ii) how increase in nutrient uptake  affects the dry matter production and yields of maize and soybean under different planting patterns; (iii) to suggest an appropriate planting pattern which is better in utilizing the nutrients  and resources with respect to competition and productivity.In previous studies, scientists have mainly focused on the morphological characteristics and seed yield of intercrop species under different planting (plant density and row spacing) arrangementsR. However, the planting pattern, SI (17244 kg ha−1) and SII (4983.1 kg ha−1), and location Lezhi (18431 kg ha−1) and Renshou (5515.2 kg ha−1) produced the highest dry matter of maize and soybean, respectively. Furthermore, the DMA of maize showed the trend planting pattern, SM > SI > SII > and locations, Lezhi > Renshou > Yaan, and in soybean it showed planting pattern, SS > SII > SI > and locations, Renshou > Lezhi > Yaan, suggesting that increasing the distance between maize and soybean rows  under MSR significantly increased the DMA of soybean by reducing the negative effects of maize shade on soybean growth. For instance, for treatment SII, it increased the DMA of soybean by 61% as compared to SI treatment. Moreover, the different planting patterns and locations significantly changed the pattern of dry matter partitioning among different plant parts of maize  was found under SII, while minimum (999.9 kg ha−1) was noted in SI. Among the locations, highest soybean seed dry matter (1689.3 kg ha−1) was recorded at Renshou, whereas, lowest (1252.4 kg ha−1) was measured at Yaan in both years. In addition, the planting pattern, SI (7485.7 kg ha−1) and location, Lezhi (7593.2 kg ha−1) produced the highest maize seed dry matter, while lowest seed dry matter of maize was found under SII (7150.1 kg ha−1) at Yaan (7007.1 kg ha−1). On average, treatment SII significantly increased the dry matter allocation to soybean seed by 41, 46, and 40% in 2012 and 38, 36, and 27% in 2013 at Yaan, Renshou, and Lezhi, respectively than those of under SI treatment.The mean values for dry matter accumulation (DMA) showed that different planting patterns and locations had significant impact on DMA of maize and soybean Table . In our ize Fig.  and soybize Fig. . At matu−1 under SM and SS, 208.8 and 126.4 kg ha−1 at Lezhi and Renshou, respectively in both study years , straw (79.1 and 44.5 kg ha−1) and seed (104.6 and 122.9 kg ha−1) N concentration of maize and soybean in SM and SS, respectively was higher than the corresponding values under SI and SII treatments. In all treatments , seed and straw of maize  and straw (97.9 kg ha−1) N concentration was measured at Renshou and Lezhi, respectively, whereas mean lowest N concentration in root (5.7 kg ha−1) and straw (64.8 kg ha−1) was observed at Yaan in both years. Furthermore, the average maximum root (4.7 kg ha−1) and seed (126.4 kg ha−1), and straw (54.7 kg ha−1) N concentration in soybean were determined at Renshou and Yaan, respectively as compared to other locations. Overall, soybean plants accumulated 68, 69, and 64% higher N in SII than SI at Yaan, Renshou, and Lezhi, respectively.Different planting patterns and locations significantly (p ≤ 0.05) affected the total nitrogen accumulation (TNA) in maize and soybean. The mean maximum TNA was 190.9 and 141.7 kg haize Fig.  and soybize Fig. , respect−1), and straw (10.5 kg ha−1) and seed (23.1 kg ha−1) P concentration of maize were significantly (p ≤ 0.05) higher in SM and SI, respectively than other treatments , straw (11.3 kg ha−1) and seed (18.9 kg ha−1) was found considerably (p ≤ 0.05) higher in SS than those of under SI and SII in MSR  and seed (24.3 kg ha−1), and straw (10.8 kg ha−1) of maize was obtained at Renshou and Yaan, respectively, and the average maximum root (0.9 kg ha−1), straw (8.7 kg ha−1), and seed (16.7 kg ha−1) P concentration of soybean was measured at Renshou, Yaan, and Lezhi, respectively. In addition, the different planting patterns significantly (p ≤ 0.05) affected the total P accumulation in maize and soybean at all locations. The average highest total phosphorus accumulation (TPA) of maize and soybean respectively was 34.5 kg ha−1 in SI and 31.5 kg ha−1 in SS, 35.2 kg ha−1 at Renshou and 25.2 kg ha−1 at Lezhi in both study years  and straw (169.9 kg ha−1), and seed (33.5 kg ha−1) in SM and SI, respectively , straw (61.1 kg ha−1) and seed (35.2 kg ha−1) K concentrations of soybean were recorded under SS , straw (174.4 kg ha−1 and 55.5 kg ha−1), and seed (33.1 kg ha−1 and 30.1 kg ha−1) of maize and soybean, respectively was measured at Renshou. Overall, soybean plants under SII accumulated 56, 65 and 85% higher K in root, straw, and seed, respectively as compared to SI. Furthermore, all the planting patterns and locations significantly (p ≤ 0.05) affected the total K accumulation in maize and soybean, and the mean highest total potassium accumulation (TKA) was 222.6 and 100.2 kg ha−1 under SM and SS, 232.5 and 88.5 kg ha−1 at Renshou, in both study years  and SII (269.7 kg ha−1), and location Lezhi (447.2 kg ha−1) and Renshou (285.2 kg ha−1) accumulated the highest nutrients by maize .The average values for total nutrient uptake (TNU) exhibited that different planting patterns and locations had significant impact on TNU of maize and soybean Fig. . In thisize Fig.  and soybize Fig. , respect−1) and harvest index (49.6%) of maize were found at Lezhi and Yaan, respectively. In addition, with the increased distance between maize rows (SI), the partial LER value of maize also increased, and the maximum LERm (1.0) was noticed at Renshou. Soybean seed yield depicted the significant (p ≤ 0.05) response to all treatments and locations. The mean maximum seed yield of soybean was noted in SS (2012.0 kg ha−1) followed by SII (1721.5 kg ha−1) and SI (992.4 kg ha−1). However, among locations, the average highest seed yields of soybean were recorded for Renshou (1640.0 kg ha−1) and lowest for Yaan (1492.9 kg ha−1) in both years. Planting pattern also showed the significant (p ≤ 0.05) effect on HI of soybean, and the mean maximum (35%) HI was measured in SII followed by SI (33%) and SS (30%). The average value of HI for locations revealed that Yaan (34%) had produced the highest value followed by Lezhi (33%) and Renshou (32%). Furthermore, with the increased distance for planting soybean between maize and soybean rows (SII) under MSR, the maximum partial LER (0.88) and CR (0.92) values of soybean were found in SII, among locations the highest LERs (0.80) and CRs (0.81) values were noticed at Renshou and Yaan, respectively. Overall, the LER under different planting patterns and locations ranged from 1.51 to 1.84, and SII produced the highest LER (1.81) at Renshou in both study years.Different planting patterns and locations led to significant (p ≤ 0.05) differences in seed yields, harvest index and partial land equivalent ratio (LER) of maize and soybean Table . By aver30. In our experiment, different planting patterns exhibited a significant effect on the DMA of maize and soybean, and highest DMA of maize and soybean was recorded in SI and SII, respectively that was probably due to the optimum planting space, light availability and nutrients availability for maize and soybean. Our findings are similar to the previously published literature in which they concluded that DMA in maize and soybean under MSR significantly affected by changing the planting pattern and light interception11. Importantly, the SII planting pattern significantly increased the soybean DMA by 61, 57, and 57% and decreased the maize DMA by 3, 3, and 5% at Yaan, Renshou, and Lezhi, respectively, maize showed small decrease in DMA as compared to a gain of dry matter by soybean plants  and yield. Since different planting patterns significantly influence the light environment under intercropping systems, and mutual shading considerably affects the productivity of intercrops because sun-light plays a vital role in increasing the photosynthetic rate and biomass yieldan Table , could e33. However, less consideration has been given to major nutrient behavior among plant parts of intercrop species in their intercropping mixtures. In this study, nitrogen, phosphorus, and potassium concentrations among plant parts of maize and soybean varied with the planting patterns and locations, but their accumulation patterns were different in all treatments. Different planting patterns significantly affected the root, straw, and seed nitrogen concentration of maize and soybean, which might be linked to the high and low nitrogen requirement of maize and soybean, respectively in all three plant parts in different planting conditions35. In cereal-legume intercropping systems, it is possible that the maize  accumulated nitrogen that was released by soybean (legume) root and root nodule-system, resulting from efficient utilization of nutrients39. Additionally, in our experiment nitrogen concentration in roots, straw, and seeds of maize and soybean in sole maize (SM) and sole soybean (SS) was higher as compared to those plants under SI and SII at all locations, suggesting that intercropping mixtures reduced the nitrogen accumulation amount in root, straw, and seed  reported the decreased nitrogen and phosphorus concentration in maize under relay intercropping system than sole cropping system27. However, the nitrogen concentration differences between relay-cropped maize and sole cropped maize were smaller as compared to soybean  and soil and soil-rhizosphere phosphorus concentration increased44. Unfortunately, in this experiment, we have not determined the phosphatase activity and soil-rhizosphere pH in the roots and soil. Interestingly, the differences of total phosphorus uptake  as compared to SI where soybean plants experienced severe shading conditions which showed the direct relationship between light intensity and phosphorus uptake because several scientists have proved that shading conditions significantly reduced the phosphorus uptake ability in crops47. Therefore, higher accumulation of phosphorus in maize and soybean is might be due the improved light conditions and growing space for intercrop species under SII (narrow wide row planting pattern) than SI.Phosphorus concentrations among different plant parts of soybean plants under MSnts Fig. , howeversed Fig.  as the dR in response to different planting patterns at different locations  between MSR and sole cropping system by soybean plants , a high nitrogen content in leaf may provide adaptation to water scarce conditions and competition by exploiting the greater sunlight availability50.In this paper, we also measured the differences of potassium uptake and distribution in different plant parts of maize and soybean under MSons Fig. . The potnts Fig.  was largnts Fig. . Overall53. Therefore, these experiments were carried out to determine the effects of different planting patterns on total nutrient uptake of maize and soybean at different locations. The findings of the present study reported here demonstrate that total nutrient uptake (TNU) in soybean was increased by increasing the distance between maize and soybean rows under MSR. Specifically, soybean plants grown under SII accumulated 18% higher TNU than those plants in SI while SII decreased the TNU of maize by 4% than SI in both years  in SII enhances the photosynthesis, carbon fixation, and plant growth rate of soybean plants. To maintain these processes at optimum level soybean plants uptake higher amount of N, P, and K in SII than SI. Results were similar with previous findings in which scientists reported that nutrients uptake increased in lupin  with increasing the light intensity54. Moreover, higher light  intensity improved the distribution pattern of major nutrients in crops which significantly increased the dry matter production47.Previously, it has been reported that nutrient uptake and nutrient use efficiency were greatly reduced by decrease in light intensityars Fig. . This enR, SM, and SS at different locations was evaluated, and the results of our study revealed that the total yield of maize and soybean were significantly (p ≤ 0.05) increased with optimum strip width arrangement SII at all locations  improved the seed yield of soybean at all locations due to the substantial increase in DMA and nutrient uptake by maintaining the maize seed yield. In the past study, researchers have reported that balanced fertilization and optimum uptake of major nutrient significantly increased the intercrop yields under MSR24. Total LER values were always higher than one in both planting patterns in MSR at all locations, which exhibits the yield benefit of MSR over sole cropping due to the better utilization of land and environmental resources for crop growth and development12. Particularly, the mean values of LER under SI and SII were 1.84 and 1.51  who reported the MSR advantage with narrow wide row planting pattern11. Similarly, Liu et al., (2017) reported higher LER values 1.3–1.4 in narrow wide row intercropping system of maize and soybean59. In addition, less distance (52 cm) between maize and soybean rows negatively affected the light interception at the soybean canopy61, therefore it is an effective method to ameliorate the negative effects of maize shade by increasing the distance  between maize and soybean rows for planting soybean, which resulted in higher light interception9, dry matter14 and seed yield production of soybean and ultimately the total LER of relay-intercropping system. In addition, the partial values of CR clearly showed maize as the dominant crop species under MSR  were greater than cereal (rye)64, which was the different trend which we observed as cereal (maize) was more aggressive and competitive than legume (soybean).The effect of different planting patterns on yield of maize and soybean under MSns Table . This in51 Table , respectR was a result of higher dry matter accumulation. In SII, 40:160 optimum strip width for growing maize and soybean, it is the best planting pattern to minimize the shading effect of maize on soybean at critical stages during the co-growth period i.e. vegetative stage and the start of flowering stage. Therefore, planting pattern SII can be adopted in MSR to avoid sever shading of maize at flowering stage. In MSR wide distance between maize and soybean rows potentially increased the land equivalent ratio (1.84) which was the result of adequate maize  and soybean  nutrient acquisition in SII , we can achieve higher seed yield and economic return under MSR. Furthermore, planning the long-term field observations to understand the impact of various planting pattern and intercrop combinations on the nutrients uptake of intercrop species is an important direction for future research.In our experiment, relay-cropped maize and soybean under different planting patterns had accumulated lower nutrient  than those of under SM and SS. However, the differences in nutrient uptake and distribution among different plant parts for soybean between relay-cropped and sole cropped plants were larger than maize plants under SI and SII planting patterns. Furthermore, the high seed yield of soybean under SII in MS−1, total N = 1.6 g kg−1, total P = 1.28 g kg−1, total K = 16.3 g kg−1, available N = 317.1 mg kg−1, available P = 42.2 mg kg−1, and available K = 382.1 mg kg−1, at Renshou pH = 6.8, organic matter = 17.3 g kg−1, total N = 0.9 g kg−1, total P = 0.5 g kg−1, total K = 14.3 g kg−1, available N = 77.4 mg kg−1, available P = 10.2 mg kg−1, and available K = 197.1 mg kg−1, and at Lezhi were pH = 6.8, organic matter = 11.1 g kg−1, total N = 1.0 g kg−1, total P = 1.4 g kg−1, total K = 17.9 g kg−1, available N = 165.1 mg kg−1, available P = 4.9 mg kg−1, and available K = 391.1 mg kg−1, in the 0–20 cm soil layer.Three experimental sites, Yaan, Renshou, and Lezhi, with different altitude levels and rainfall characteristics, were selected for this experiment. The field experiment did not include any protected or endangered species and no permissions were needed for the selected locations. All the experiments were conducted by following the institutional rules and regulations of Sichuan Agricultural University, China. Yaan  has a humid climatic condition with mean annual temperature of 16.2 °C and rainfall 1200 mm. The subtropical and humid site Renshou , has average rainfall of 1009.4 mm and temperature 17.4 °C. Lezhi  has monsoon humid, with mean rainfall of 949.4 mm and temperature of 16.7 °C. Weather data which includes monthly rainfall, average temperature, humidity and wind speed during the planting seasons from 2012 to 2013 are shown in Table −2 for maize and soybean, respectively in relay-intercropping system, and similar planting density was kept in sole cropping system of maize and soybean. The maize crop was sown in the second week of April in 2012 and 2013, and harvested in the first week of August 2012 and 2013. Soybean was sown on in the second week of June 2012 and 2013 and harvested in the last week of October 2012 and 2013. All plots were treated with basal fertilizer. Basal nitrogen (N) at 135 kg ha−1 as urea, phosphorus (P) at 40 kg ha−1 as calcium superphosphate, and potassium (K) at 10 kg ha−1 as potassium sulfate were applied at the time of sowing in intercropped and sole-cropped maize. At the V6 stage of maize, the second dose of N was applied at 75 kg ha−1 as urea in all plots. The N, P, and K at 75, 40, and 4 kg ha−1 as urea, calcium superphosphate, and potassium sulfate, respectively were basally applied for soybean at the time of soybean sowing. Other farming measures were used according to the farmer’s practices.The maize genotype ‘Chuandan-418 (semi-compact)’ and the soybean genotype ‘Nandou-12 (shade resistant)’ were selected for the experiments. This field study consisted of four different planting pattern arrangements Fig. , describTen consecutive maize and soybean plants, excluding the five border plants, were destructively sampled from all experimental blocks at maturity stage during both years and locations for the determination of total dry matter accumulation (DMA) and partitioning in different plant parts of maize and soybean. Then all the sampled plants were divided into different plant parts of maize , and seed) and soybean , and seed) placed in oven for one hour at 80 °C to kill the fresh-tissues and then dried at 65 °C to obtain constant weight before weighing of each plant part of maize and soybean for total dry matter accumulation and partitioning analysis.3, the phosphorus (P) content of maize and soybean plant samples was determined by using the vanadomolybdate procedure55, and the potassium (K) content of maize and soybean plant samples were estimated by following the previously described method65. The N, P, and K content in each organ of maize and soybean plants were measured by multiplying the total dry matter of each plant organ with the N, P, and K content and calculated in a kg ha−13. The total N accumulation (TNA), total P accumulation (TPA), and total K accumulation (TKA) were calculated from the summation of the nitrogen, phosphorus, and potassium in all plant parts56. Additionally, total nutrient uptake (TNU) was measured from the summation of the TNA, TPA, and TKA in maize and soybean.At maturity of maize and soybean, plant samples were collected from the central rows of each experimental block and location, divided into root, straw, and seed of maize and soybean. Then all the maize and soybean samples were dried in the oven at 80 °C for 96 hours to attain constant dry weights. Then dry matter of each plant part was ground using a ring-mill through hundred-mesh and the nitrogen (N) content of maize and soybean plant samples was measured by using the Kjeldahl method−2 plants of maize and soybean were manually harvested from the central rows of each experimental block using sickle at ground level. Then these sampled plants were dried for ten days. The dried maize and soybean plants were threshed manually and weighed to determine the intercropped and sole cropped maize and soybean seed yields of every plant and then converted into kg ha−1. In addition, harvest index (HI) was estimated by dividing the seed weight per plant by the dry matter weight of each plant at physiological maturity. The relay-intercropping advantage and competition effects between two different crop species grown in mixture were determined by using several competition parameters as follows: the LER was applied as the standard for intercropping advantage because both maize and soybean were the preferred crop species in relay-intercropping system. One is considered as the crucial LER value because if the value of LER is greater than one then it means that intercropping system favors the crop growth and yield of intercropped species, while if LER value is less than one then that intercropping system negatively reduces the growth and yield of intercropped species66. LER was determined as:In this experiment, at the time of maturity, four m67. In addition, Microsoft Excel program was used for the graphical presentation of data using standard error (±SE).All the measured data for all parameter was analyzed using Statistix 8.1. An ANOVA  technique was used to confirm the overall significance of data. The least significance difference (LSD) test was applied to compare the means at 5 percent probability level


Cocirculation of varying influenza types, strains, and lineages allows coinfection and intra‐season sequential infection, although a same‐strain sequential infection has not been previously described. This case report describes the first known case of sequential laboratory‐confirmed influenza A (H3N2) infections in a child within one season. Briefly, since January 5, 2015, a longitudinal school‐based influenza surveillance study has been ongoing at the Oregon School District (OSD) in South Central Wisconsin. ORCHARDS (ORegon CHild Absenteeism due to Respiratory Disease Study) is designed to assess the utility of school absenteeism monitoring for early influenza outbreak detection by comparing illness‐related absenteeism with medically attended influenza in surrounding communities. Parents of children with influenza‐like illness (ILI) symptoms voluntarily call a study hotline; students are screened for ILI criteria and, if eligible, receive a home visit from a research team member. Research staff collect nasal and naso/oropharyngeal (NP/OP) specimens along with symptom and epidemiologic data. Specimens are tested by rapid influenza diagnostic testing  within 2 hours of the home visit.All specimens are then tested for influenza by rRT‐PCR (IVD CDC Human Influenza Virus RT‐PCR Diagnostic Panel) and other respiratory viruses (Luminex NxTAG Respiratory Pathogen Panel: RPP) at the Wisconsin State Laboratory of Hygiene (WSLH). The WSLH undergoes a rigorous validation process for diagnostic tests, including rRT‐PCR, in accordance with federal CLIA licensing standards. Unidirectional workflow, preparing aliquots of one specimen at a time, dedicated specimen processing equipment and laboratory area, and decontamination of biosafety cabinet (BSC) and laboratory benches after each use ensure cross‐contamination risks are minimized.Beginning in September 2016, families of ORCHARDS subjects were invited to participate in a home transmission substudy, for which nasal swabs are obtained from family members the day of the visit (day 0) and 7 days later, along with a day 7 nasal swab from the ORCHARDS subject. All specimens from household members are tested for influenza using rRT‐PCR as above.Exceptionally for this case study, next‐generation genome sequencing was performed at WSLH using the Illumina MiSeq™ platform. Sequence reads were trimmed using Trimmomatic with a 4‐bp sliding window quality score cutoff of Q30.The University of Wisconsin Institutional Review Board (IRB) (2013‐1357‐CP022) approved the study protocol, and the Centers for Disease Control and Prevention's Human Research Protection Office reviewed and approved deferral to the University of Wisconsin's IRB.33.1On January 31, 2018, a usually healthy, 9‐year old, non‐Hispanic white female was screened for ORCHARDS eligibility , chills, cough, wheezing, nasal congestion, headache, malaise, myalgia, anorexia, burning eyes, and dizziness. Again, there was no recent travel, exposure to farm animals, or identified sick contact preceding onset. The subject's RIDT result was positive for influenza A, and rRT‐PCR results confirmed the presence of influenza A (H3N2) (Ct = 26.48). At follow‐up, she reported 2 days of absenteeism and continued fever, chills, cough, rhinorrhea, malaise, myalgia, headache, and anorexia. There had been no medical follow‐up or antiviral treatment. The rRT‐PCR results from the subject's day 7 swab continued to show influenza A (H3N2) (Ct = 33.30). No other family members reported illness on day 0 or day 7. All family members tested negative for influenza by rRT‐PCR on day 0 and day 7.3.3® genetic sequence database and given accession numbers .The subject's initial specimen from each episode, collected on January 31 and March 9, 2018, was prepared for full genomic sequencing at the WSLH. Both A (H3N2) strains were of the hemagglutinin gene clade 3C2.a2. After WSLH performed a consensus sequence comparison between the two viruses collected from the subject, three single nucleotide polymorphisms (SNPs) were identified within the coding region of the HA protein. Only one SNP was non‐synonymous and resulted in an isoleucine to leucine change at position 67 (in HA1). This particular polymorphism was near 100% frequency in the sequence reads and was four amino acids away from an established antibody epitope site (Site E).4This is the first known case of two sequential, within‐season infections with influenza A (H3N2). Moreover, this case occurred within an immunocompetent and fully vaccinated child. The period between ILI episodes (37 days), the absence of influenza by PCR at 7‐day follow‐up of the first episode, the resolution of symptoms, and the development of a new ILI support two distinct episodes of influenza infection.In the first episode, coinfection with coronavirus HKU1 was documented as well as a high Ct value for influenza A (H3N2) and a falsely negative RIDT. This may imply a relatively low viral load and possible immune interference induced by coinfection with coronavirus.In the second episode, the subject's ILI was more significant, the RIDT was positive, and her initial specimen manifested a much lower Ct value, implying a higher viral load. This occurred despite vaccine receipt and prior infection with the same influenza subtype. The timing of the first episode, relative to the second, was such that neutralizing antibodies should have been present.It is known that influenza remains under‐detected by established surveillance systems, which rely on sentinel‐based reporting of medically attended cases.The authors have no conflicts of interest to declare.


To evaluate choroidal vascularity index (CVI) in eyes with internal carotid artery (ICA) stenosis using binarization tool in enhanced depth images scanned by spectral domain optical coherence tomography.A retrospective cohort study was conducted in 142 patients with ICA stenosis, and 20 normal control subjects matched with the age, sex, etc. According to the diagnostic criteria, the participants are divided into a normal control group (20 cases), a mild stenosis group (64 cases), a moderate stenosis group (61 cases), and a severe stenosis group (17 cases). Enhanced depth imaging optical coherence tomography (EDIOCT) was performed to scan macular fovea, which was separated into a luminal area and a stromal area using image binarization. The choroidal vascularity index (CVI) is luminal area divided by total choroidal area.P < 0.05). Moreover, the CVI in moderate stenosis group and severe stenosis group were significantly lower compared with the normal control group (P < 0.001). When CVI = 65.16% was set as the cut-off value, all 162 subjects could be divided into two groups, CVI ≤ 65.16 (n = 83) and CVI > 65.16% (n = 79). The proportions of mild stenosis, moderate stenosis, and severe stenosis in CVI ≤ 65 .16 group and CVI > 65.16% group were statistically significant (P < 0.001).There was no statistical difference in age or sex among groups. Subfoveal choroidal thickness (SFCT) in the severe stenosis group was significantly lower than that in the normal group (CVI may be a useful indicator for early diagnosis and monitoring of choroidal vascular changes in ICA stenosis. Internal carotid artery (ICA) stenosis or obstruction may lead to severe ischemic cerebrovascular disease, and is also an important cause of ischemic eye disease, which not only affects the patient’s vision, but even causes physical disability and sudden death. When ICA stenosis is greater than 50%, the incidence of ocular symptoms increases , 2. HemoWith the emergence of new optical coherence tomography (OCT) techniques, the study of choroidal structure has made great improvement , 6, 7. SThe application of choroidal structure image binarization to investigate the structure of the choroid and its changes in different disease models have been investigated in recent years. A number of studies have demonstrated that choroidal structure image binarization are rarely affected by physiological variables and are capable of providing choroidal vascular information –19. In oThis retrospective cohort study was conducted in the Department of Ophthalmology, Beijing Friendship Hospital from January 2016 to December 2017. A total of 179 patients diagnosed with ICA stenosis/obstruction were enrolled and divided into four groups according to the North American Symptomatic Carotid Endarterectomy Trial (NASCET) criteria [NASCET stenosis classification method was used to calculate the degree of stenosis according to the following formula: stenosis degree% = *100%. The diameter (N) of the narrow part and the diameter (D) of the distal normal vessels were measured. The stenosis rate r = (1-N/D) × 100%. Stenosis was graded: no stenosis (stenosis rate 0), mild stenosis (stenosis rate < 29%), moderate stenosis (stenosis rate 30–69%), severe stenosis (stenosis rate 70–99%) and complete occlusion.ICA stenosis or plaque formation was confirmed by head- and neck- CT. The ICA patients with following ocular characteristics were excluded: diopter > ± 3.0 , active intraocular inflammation and/or infection, or any type of intraocular surgery (except cataract surgery) and retinal neovascularization caused by diabetic retinopathy or other macular disease. Additionally, the patients with the systemic diseases were also excluded: patients with peripheral vascular disease, limb paralysis, severe eye disease history ; acute myocardial infarction, stroke and infectious inflammatory disease in the first few weeks of the study; patients with persistent heart rate abnormalities and chronic heart failure; pulmonary hypertension and patients with acute or chronic infectious diseases or malignant tumors; obvious refractive interstitial opacity or abnormality affecting OCT measurement, such as obvious cataract, vitreous opacity, vitreous hemorrhage, vitreous cavity silicone oil filling state, etc.; patients who have undergone internal carotid artery surgery.A total of 159 patients and 20 control subjects were included in the data collection. The data included the following contents: general information ; health status ; daily life habits ; eating habits ; and family history .Ocular data were collected from all enrolled patients, including best corrected visual acuity (BCVA), non-contact intraocular pressure measurement, slit-lamp ophthalmoscopy, color fundus photography, enhanced depth imaging optical coherence tomography (EDIOCT) . The OCT image acquisition time was 9–12 AM daily.The artery occlusion was selected on the heavier side. One eye with clear choroidal image was selected for analysis in normal subjects and the patients with the similar occlusion in both sides. All data were collected by the same technician (Wang Wei). After the EDIOCT, the best image was displayed on a computer screen and independently evaluated by three physicians. When two or more graders determine that the foveal choroid image was clearly identifiable, the image was considered acceptable and used for analysis. For pictures with unclear choroidal image, two physicians explored the boundary. If there were significant discrepancies between the two, the third physician was inquired. Finally, 17 images were excluded, and the remaining 162 images were obtained from the four groups, including no carotid artery stenosis group (20 cases), mild stenosis group (64 cases), moderate stenosis group (61 cases), and severe stenosis group (17 cases).Subfoveal choroidal thickness (SFCT) was measured in the fovea of the macula. It was the distance from the outer surface of the retinal pigment epithelial layer to the choroid and sclera interface below the fovea Fig. . The nasUsing the Auto Local Threshold (Niblack Method) and polygon selection tool, the choroidal region was marked with bruch membrane as the upper boundary and choroidal-scleral interface as the lower boundary, with the central fovea as the center, and the distances between nasal and temporal sides were 750 μm. The total choroidal area (TCA) was calculated, and the image was converted into 8-bit. The Auto Local Threshold was used to perform image binarization. In the binarized image, dark pixels represented the vascular cavity, and white pixels represented stromal area. After converting the image into RGB  color, the dark pixels were selected using the color threshold tool to calculate the area of the dark pixels of the luminal area (LA). Stromal area (SA) was calculated by subtracting LA from the TCA. The ratio of LA to TCA is defined as CVI Fig. .Fig. 2EP values were less than 0.05.The data was analyzed and processed using SPSS 19.0 software. Countable data were analyzed by χ2 test. One-way ANOVA was used to compare the data among groups, and the data were expressed as mean ± standard deviation (SD). The difference was statistically significant when P = 0.734). There was no significant difference in the mean age among the four groups (P = 0.892). There were no differences regarding the incidence rates of diabetes, cerebrovascular disease coronary heart disease in four groups. Therefore, we ignored the impact of these factors on the final analysis  and 8 females (40%); 43 males (67.2%) and 21 females (32.8%) in the mild carotid stenosis group; there were 42 males (68.9%) and 17 females (30.4%) in the moderate stenosis group. There were 10 males (58.8%) and 7 females (41.2%) in the severe stenosis group. The gender distribution of the four groups was not statistically significant (p = 0.355 and p = 0.945). The significant difference was only observed between control and severe stenosis group (P < 0.05) , 259.25 ± 30.07 μm (95% CI: 253.73–266.76), 252.42 ± 30.78 μm (95% CI: 244.53–260.30), and 230.32 ± 30.80 μm (95% CI: 214.4–246.16), respectively. There was no significant difference between the ICA mild stenosis group and the ICA moderate stenosis group compared with the normal control group (P = 0.219). When compared with normal control group, the values in ICA moderate stenosis group and ICA severe stenosis group were statistically reduced (P < 0.001). The receiver operating characteristic (ROC) curve was shown in Fig. The CVI of the normal control group was 66.84 ± 1.45% (95% CI: 66.16–67.51%), 66.39 ± 1.8% (95% CI: 65.94–66.83%) in the ICA mild stenosis group, 64.42 ± 0.9% (95% CI: 64.19–64.65%) in the ICA moderate stenosis group and 62.76 ± 1.01% (95% CI: 62.24–63.28%) in the ICA severe stenosis group (Table n = 83) and CVI > 65.16% (n = 79) (Table P = 0.912). There was no significant difference in the mean age between the two groups (P = 0.604). There was also no significant difference in the proportion of hypertension, diabetes, cerebrovascular disease and coronary heart disease between the two groups. Therefore, we ignored the impact of these factors on the final analysis.We used CVI = 65.16% as the cut-off value. All 162 participants were divided into two groups, CVI ≤ 65.16% n = 8 and CVI P < 0.001) , while the SFCT in the mild stenosis group and the moderate stenosis group was comparable with the normal group. The CVI of the severe stenosis group was significantly lower than that of the normal group, indicating that the choroidal thickness did not change significantly during moderate stenosis, but the ratio of vascular components in the choroid had changed greatly. The area of LA was larger than that of the normal group. However, the area of SA was significantly lower than that of the normal group. The blood supply to the eye tissue mainly comes from the ophthalmic artery. The reduction of blood supply to the ophthalmic artery will inevitably affect the retina and choroid. Choroidal blood flow accounts for 90% of ocular blood flow, and has important physiological effects.We hope to find a diagnostic indicator to detect changes in ocular blood flow in ICA patients. A CVI of 65.16% was set as a cut-off value after analyzing the ROC curve and all cases were divided into two groups. The composition between the two groups was significantly different. CVI > 65.16% group mainly included normal control group, ICA mild stenosis cases and a small portion of moderate stenosis cases, but without severe stenosis cases. CVI ≤ 65.16% group mainly included patients with moderate stenosis group and all patients with severe stenosis group. A small number of patients with mild stenosis were included in CVI ≤ 65.16% group, which also confirmed our hypothesis that when the ICA stenosis is aggravated, the eye presents a chronic ischemic state, which not only affects the choroidal capillaries, but also reduces the diameter. It affects the small blood vessels in the Haller’s layer, and the inner layer of the Sattler layer, resulting in a decrease in the proportion of the choroidal vasculature.Our study has several limitations. Firstly, all the observers enrolled in this study were from hospital patients, and the prevalence of various cardiovascular diseases such as cardiovascular and cerebrovascular diseases was slightly higher than that of the normal population, although without significant difference when compared with controls. In addition, bias might be caused by the small number of patients in the severe stenosis group. The future study will expand the enrollment group, especially those in the severe stenosis group. Secondly, we only measured a single scan of the 1500-μm region centered on the fovea, and a larger region or macular volume scan may provide us with better choroidal vascular index information.In summary, the use of choroidal image binarization to analyze choroidal structural changes in patients with ICA stenosis provides us with a possibility to correlate the choroidal vasculature with the large vascular system of the body. The patients with CVI ≤ 65% have a greater possibility of serious occurrence.


Attentional biases are a relatively robust phenomenon among clinical populations but less pronounced in healthy participants. However, regarding the components of attentional biases and the directions of attention allocation, there are several inconsistencies in the literature. The present study examined whether these inconsistencies can be traced back to previous experiences of relational peer victimization in clinical populations.n = 30) and healthy controls (n = 31). Additionally, the sample was divided into two subgroups according to the participants’ reports of previous relational peer victimization . Attentional biases were measured by the Emotional Stroop task and a dot-probe task.Participants were subjects with a diagnosed psychiatric disorder (In both samples, peer victimized participants showed delayed response times when color-naming negative and positive compared to neutral adjectives in the Emotional Stroop task. Likewise, the dot-probe task indicated attentional avoidance of both negative and positive words in peer victimized participants with and without a psychiatric disorder. Interestingly, presence of a psychiatric disorder did not have a significant effect on attentional biases.Both tasks could detect that attentional processes were linked to the experience of peer victimization rather than to the current diagnostic status of the participants. Attentional avoidance of emotional stimuli may prevent victimized individuals from responding adequately to environmental stimuli, which may increase the risk for the development of psychopathology. A large body of research has demonstrated that attentional biases are a relatively robust phenomenon among anxious populations, but less pronounced and consistent in non-anxious subjects –5. GenerThe most commonly used task to measure attentional bias is the modified or Emotional Stroop task . In thisFrom a clinical point of view, attentional biases are of interest because they are most likely relevant in the development and maintenance of psychiatric disorders , 11. ForAlthough the majority of studies demonstrated a difficulty in disengagement among anxious individuals , 31, 32,With respect to attentional biases in other psychiatric disorders, the current body of literature is peppered with inconsistencies as well. A meta-analysis of 29 empirical studies examining depressive patients demonstrated an attentional bias towards negative information in this population . While dAccordingly, it appears that trait anxiety and psychopathology are associated with attentional biases. However, this association seems to be moderated by both parameters within the paradigms used to detect biases   and indiTo date, most studies examined ACEs including low maternal care, parental conflicts , 63, or The aim of the present study was to address inconsistencies in the existing literature about attentional biases in clinical samples. Here, a large body of studies presenting an attentional bias towards negative stimuli , 30, 46 The current study sought to address this underrepresentation by examining the influence of peer victimization on attentional biases as measured by the Emotional Stroop task and the dot-probe task in two samples consisting of subjects with either a diagnosed psychiatric disorder or healthy controls. With respect to the existing literature, we hypothesized that subjects with a diagnosed psychiatric disorder would show an attentional bias towards negative compared to neutral adjectives. Furthermore, we assumed that the attentional bias would be more pronounced in subjects with a history of peer victimization irrespective of their current diagnostic status. When comparing positive to neutral adjectives, we did not expect to find any attentional biases.Due to the study’s aims, recruitment of participants was two-pronged. The clinical sample was recruited through the Hans-Peter-Kitzig-Institut , a regional rehabilitation hospital for patients with psychiatric disorders. The healthy control sample was recruited through online advertisements in student newsgroups and bulletins at the campus of Bielefeld University. Advertisements informed that the study examined the association of personality traits, life experiences, and attentional processes.The total sample consisted of 61 participants, . Out of the whole sample, 30 individuals (49.2%) represented the clinical sample. Exclusion criteria for the clinical sample included (a) evidence of a current substance abuse or dependence, (b) evidence of current active-phase symptoms of psychosis as delusions, and hallucinations, and (c) evidence of acute suicide intention or ideation. Number and types of diagnoses of the clinical sample are presented in Table Information about the diagnostic status  of the clinical sample was obtained from the participants’ health records of the Hans-Peter-Kitzig Institut. Diagnostic status of the control sample was assessed using the German Version of the Mini International Neuropsychiatric Interview (M.I.N.I.) –85. The The stimulus set consisted of 180 adjectives  and was derived from prior studies on word processing –88. In tThe Emotional Stroop task consisted of 240 trials. In total, 80 negative, 80 neutral, and 80 positive words were shown, in each case 20 words were colored in red, 20 in blue, 20 in green, and 20 were colored in yellow on a black background. Each single word was presented four times. Stimuli were shown throughout until the participants responded. After an intertrial interval of 200 ms the next stimulus was presented. Participants’ task was to identify the color of the presented words as quickly and as accurately as possible. Participants indicated their response by pressing buttons on a keyboard with the index and middle fingers of both hands. In order to reassure that the participants were able to assign the colors to the right buttons, the assignment of buttons and colors was presented on the screen throughout the experiment. The assignment of buttons was counterbalanced across participants. The order of words, word valences, and colors was randomised. We used the software package Inquisit 4.0.3  to deliver stimuli and record responses and reaction times (RTs).The dot-probe task consisted of two blocks of 240 trials each, with a short break between the blocks. There were three different types of trials in the present task: negative–neutral, positive-neutral, and neutral-neutral, with negative and neutral, neutral and neutral, and positive and neutral words combined, respectively. All words were presented in black on a white background, lowercase. The word pairs were presented with one word beside the other  in the middle of the screen. The dot-probe experiment began with 12 practice trials using neutral–neutral word pairs to familiarize participants with the task. Each trial started with a black fixation cross presented in the center of a white screen for 500 ms. Then, a word pair appeared with one word beside the other for 500 ms. A gray dot emerged in one of the word locations immediately after the offset of the words. The location of the target word (left or right) and probe (left or right) was randomized for all trials. The inter-trial interval for all trials was 500 ms. Participants were instructed to respond as quickly and as accurately as possible and to indicate the location of the gray dot (left or right) by pressing either the button “E” (left) or “I” (right) on a keyboard with their index fingers of both hands. The three types of word pairs were randomly formed. Each word was presented six times (3 times on each side) for a total of 480 experimental trials. The combination and order of word pairs varied randomly for each participant. We used the software package Inquisit 4.0.3  to deliver stimuli and record responses and reaction times (RTs).Fragebogen zu belastenden Sozialerfahrungen, FBS [Adverse Social Experiences Questionnaire]) ) . The FBSr = .89) , as wellr = .89) , 89. Ther = .89) , 91–93).r = .89) , 94, symr = .89) , 96, genr = .89) –99, and r = .89) , 101. Inr = .89)  to deterM = 5.65, SD = 4.17; control sample: M = 6.29, SD = 4.26; t (57) = .58, p = .562). No participants were excluded due to higher error rates than 25%. Outliers were defined as participants presenting reaction times that deviated more than three SDs from mean reaction times and were removed from analyses. Outliers were present in both the clinical (n = 1) and the control sample (n = 1).In the Emotional Stroop task an attentional bias is indicated by greater color-naming latencies following negative/positive words in comparison with neutral words . TherefoFor the dot-probe task, attentional bias scores were calculated for two different trial types . Here, an attentional bias is indicated by either lower RTs to the probe if it emerges at the location where the participants were focusing their attention, or higher RTs to the probe when it appears in the location where the participants were not attending . ParticuTo better understand the mechanisms underlying the attentional bias, additional index scores were calculated to differentiate vigilance and difficulty to disengage . VigilanM = 10.52, SD = 10.12; control sample: M = 8.13, SD = 5.71; t (56) = 1.11, p = .273). No participants were excluded due to higher error rates than 25%. Outliers were defined as participants presenting reaction times that deviated more than three SDs from mean reaction times and were removed from analyses. Outliers were present for the clinical (n = 1) and the control sample (n = 2). Accordingly, the remaining sample for the analyses of the Emotional Stroop task consisted of 59 participants  and for the analyses of the dot-probe task of 58 participants .In line with previous studies, trials with reaction times lower than 150 ms or higher than 2000 ms were excluded from analyses , 106. Inn = 28) on the FBS [n = 33). Next, for explorative reasons the existence of absolute attentional biases was examined by applying planned t-tests for each attentional bias index score. Specifically, the bias and index scores for all four combinations of group  and peer victimization (high vs. low) were compared to zero. Additionally, because the clinical sample and the healthy control sample differed significantly on some subscales of the CTQ, all ANOVAs were carried out as analyses of covariance (ANCOVAs) with age and the CTQ sum score serving as covariates to control for the influence of childhood maltreatment within the family. As the pattern of results did not change, only ANCOVAs with age serving as the covariate are reported. For the ANCOVAs, partial eta-squared (η2) values were reported to demonstrate the size of effects such that 0.01 represents a small effect, 0.06 a medium effect, and 0.14 a large effect [.All statistical analyses were carried out using the Statistical Package for the Social Sciences 25. Because age differed significantly between the two samples, all ANOVAs were carried out with age serving as a covariate. Initially, an omnibus 2  × 2 (peer victimization: high vs. low) × 3  analysis of covariance (ANCOVA) with repeated measures on valence and age serving as a covariate was calculated for the mean RTs of the Emotional Stroop task. Similarly, an omnibus 2  × 2 (peer victimization: high vs. low) × 2  × 2 (location of the dot: congruent vs. incongruent) analysis of covariance (ANCOVA) with repeated measures on valence and age serving as a covariate was calculated for the mean RTs of the dot-probe task. Afterwards, several 2  × 2 (peer victimization: high vs. low) analyses of covariance (ANCOVAs) were conducted to examine the different hypothesized attentional biases dependent on the extent of diagnostic status and peer victimization. To date, the FBS lacks a representative norm sample and validated cutoff scores for peer victimization. In line with previous studies , 91, 92, the FBS . Particie effect .M = 24.59 years (SD = 4.97). However, mean age differed significantly between the two samples  = 2.61; p = .012). Participants’ means on the assessments and diagnoses are presented in Table The average age of the total sample was F  = 3.30; p = .041; η2 = .060. Further main or interaction effects did not reach significance  = 3.90; p = .053; η2 = .067  = .11; p = .747; η2 = .002. The interaction effect of Group x Peer victimization did also not show significant differences, F  = .19; p = .662; η2 = .004.For negative-neutral trials, the age-corrected ANOVA revealed a marginally significant main effect of peer victimization, F , 54 = 3.F  = 4.57; p = .037; η2 = .081. There was no main effect of group, F  = .06; p = .803; η2 = .001. Similarly, the interaction of Group x Peer victimization did not reach significance, F  = .37; p = .548; η2 = .007. Additionally, the Emotional Stroop Index score for the positive-neutral trials showed a significant difference from zero in healthy controls without a history of peer victimization, t (22) = − 2.56; p = .018. In the explorative analyses, all other Emotional Stroop index scores did not differ significantly from zero .For positive-neutral trials, the age-corrected ANOVA showed a significant main effect of peer victimization, F  = 5.89; p = .019; η2 = .100; group: F  = 16.21; p < .001; η2 = .234). Additionally, the interaction effects of Location x Group, F  = 4.77; p = .033; η2 = .083, and Location x Peer victimization, F  = 6.20; p = .016; η2 = .105, reached significance. Further significant main or interaction effects were not found  = 6.27; p = .015; η2 = .106, for the general attentional bias score when comparing negative to neutral words  = 2.46; p = .123; η2 = .044, and the interaction effect of Group x Peer victimization, F  = .83; p = .367; η2 = .015, were not found to be significant. The age-corrected ANOVA for the attentional bias score for positive compared to neutral words did not show any significant effects  indicating that only relative attentional biases were present.The ANOVA with age serving as a covariate revealed a significant main effect for peer victimization, p’s > .05; see Table p’s > .05; see Table p’s > .05).For the orienting index score comparing negative to neutral words, the age-corrected ANOVA did not reveal any significant effects  = 3.20; p = .080; η2 = .057. In the explorative analyses, the analyses of absolute bias scores showed no significant effects .The ANOVAs with age serving as the covariate for the analyses of the effects of peer victimization and group on the difficulty to disengage from negative or positive words showed no significant effects  in highly peer victimized participants. With respect to the results of the dot-probe task, peer victimized participants’ responding can be interpreted as attentional avoidance of emotional stimuli. Reported effect sizes of the significant effects were medium.In accordance with studies indicating attentional biases in subjects with ACEs –61 and oIt may be speculated that for subjects encountering situations of peer abuse there are initially no active coping or behavioral resources based on a fight or flight stress response which would be associated with facilitated attention towards threat. Instead, subjects experiencing peer victimization may undergo a down-regulation of behavioral and attentional processes which are reflected by avoidance of threatening stimuli. This reasoning is supported by the fact that peer victimization is associated with blunted responses to stress , 113–118However, since the pattern of results did not differ between negative and positive adjectives, the present study indicated a general emotion-avoidant, rather than threat-avoidant, attentional style in subjects with a history of peer victimization. This finding is in accordance with other reports of attentional biases in reaction to different emotional stimuli –134 as wRecently, Rudolph, Troop-Gordon, and Granger (2010) reported that anticipatory cortisol and salivary alpha amylase activation was increased in victimized children who were informed that they would be interacting with unfamiliar peers . It was Contrasting with a wealth of research , 29, 42 Addressing the fact that diagnoses in the present study were rather heterogeneous and therefore may be accompanied by a varying magnitude and direction of biases, this conclusion also refers to the finding that peer victimization influenced attentional processes even beyond the effects of trait anxiety. However, most studies indicating attentional biases in psychiatric disorders, and particularly anxiety disorders, used stimuli that were either threat-, fear- or disorder-related , 16, 49.Moreover, the present study has several additional limitations. The assessment of peer victimization was based on self-report and retrospective accounts and may be subjected to recall biases . AnalyseRecently, a growing body of literature has emphasized the role of peer victimization as a major public health concern. In school children, reports of repeated victimizations range from 10 to 20%, with periodic adversities being indicated even more frequently , 145. Th


Healthcare-associated infections (HAIs) have a major impact on public health worldwide. Particularly, hospital surfaces contaminated with bacterial pathogens are often the origin of both sporadic cases and outbreaks of HAIs. It has been demonstrated that copper surfaces reduce the microbial burden of high touch surfaces in the hospital environment. Here we report the antimicrobial characterization of a novel composite coating with embedded copper particles, named Copper Armour™.Staphylococcus aureus, Pseudomonas aeruginosa, Escherichia coli O157:H7 and Listeria monocytogenes. Additionally, its antimicrobial properties were also evaluated in a pilot study over a nine-week period at an adult intensive care unit. For this, four high touch surfaces, including bed rails, overbed table, bedside table and IV Pole, were coated with Cooper Armour™, and its microbial burden was determined over a nine-week period.The Copper Armour™ bactericidal activity was evaluated in in vitro assays against several bacterial pathogens, including Copper Armour™ coated samples showed an in vitro reduction in bacterial burden of > 99.9% compared to control samples. Moreover, pilot study results indicate that Copper Armour™ significantly reduces the level of microbial contamination on high-touch surfaces in the hospital environment, as compared with standard surfaces.Based on its antimicrobial properties, Copper Armour™ is a novel self-sanitizing coating that exhibits bactericidal activity against important human pathogens and significantly reduces the microbial burden of hospital surfaces. This composite could be used as a self-sanitizing coating to complement infection control strategies in healthcare facilities. Healthcare-associated infections (HAIs) are the most frequent adverse event threatening the life of hospitalized patients worldwide . HAIs haStaphylococcus aureus (MRSA), vancomycin-resistant Enterococcus spp. (VRE) and Clostridium difficile, are able to colonize hospital surfaces, and both spores and the vegetative form can persist on these surfaces for months , sonicated in an ultrasonic bath  for 5 min and turned to mix. Within 1 h, serial dilutions were performed in PBS and plated in duplicate on TSA. After incubation for 24–48 h at 37 ± 0.5 °C, the number of cfu was counted. The number of cfu recovered per sample was determined taking into consideration the dilution (20x), using the following equation: cfu/sample = (A x D x V) / V2, where A = average cfu per sample, counted in duplicate; D = dilution factor; V = volume of TPL solution added; and V2 = volume plated. The percentage reduction in the number of cfu for test samples as compared with the control samples was determined using the following equation: % reduction = [(a-b) / a] × 100 where, a = geometric median of the number of cfu recovered in control samples; and b = geometric median of the number of cfu recovered in the test samples.Bacterial culture media were supplemented with 5% heat-inactivated fetal calf serum  and 0.01% Triton X-100 as organic sediment load. Initial inocula  and six control samples  were evaluated per microorganism in a similar fashion to that described in Protocol 1. Samples were consecutively inoculated eight times, adding the challenging microorganism at 0, 3, 6, 9, 12, 15, 18 and 21 h. The antimicrobial efficacy was evaluated at 2, 6, 12, 18 and 24 h, corresponding to 1, 2, 4, 6 and 8 inoculations. After exposure to bacteria, 20 ml of TPL solution was added and samples were subjected to sonication in an ultrasonic bath and turned to mix. The determination of the number of cfu recovered per sample and the percent reduction was performed as described for protocol 1. Additionally, we performed the same sterility controls as previously described.The study was conducted in two patient rooms (side by side) within the adult ICU at the Hospital Clínico Universidad de Chile located in Santiago, Chile. One of the rooms was defined as the control and in the other room, considered the intervention room, surfaces were coated with Copper Armour™ Fig. . The fol2). Each dressing was placed in a 50 ml sterile polypropylene centrifuge tube. Within 2 h, three ml of PBS/LT (0.5% Tween 80 and 0.07% lecithin) were added to each centrifuge tube, vortexed for 1 min, and allowed to settle for 5 min. Subsequently, 100 μl aliquots were plated on 5% sheep blood agar to estimate the total aerobic microbial burden present on sampled surface; mannitol salt agar  to estimate the number of cfu of Staphylococcus spp.; MacConkey agar  to determine the number of cfu of Gram-negative bacilli; chromogenic agar  to estimate the number of cfu of MRSA; bile esculin agar  supplemented with vancomycin (6 μg/ml) to determine the number of cfu of VRE and Sabouraud agar  supplemented with chloramphenicol (CAF) to estimate the cfu of yeast / fungi. Plates were incubated for 24–48 h at 37 ± 0.5 °C and the number of cfu were determined. The number of cfu recovered per sample was reported as cfu/100 cm2.The sampling protocol was performed over a nine-week period, during which the first week  was dedicated to methodology adjustments. Data obtained during this week were not included in statistical analysis and are not shown. Rooms were sampled on the same day and at the same time (before morning cleaning) every week throughout the study. Surfaces were sampled in duplicate . A P-value of < 0.05 was considered statistically significant; statistical analysis was performed in GraphPad Prism version 6.00 .Data on microbial burden obtained from hospital surfaces were analyzed for normality using the Shapiro-Wilk test. As the data did not follow a normal distribution, the non-parametric Mann-Whitney U test (one-tailed) was used to determine if the microbial burden of Copper Armour™ coated surfaces was significantly lower compared to control surfaces. Additionally, differences in the frequency of microbial burden, reported as > 250 cfu/100 cmSEM analysis of samples coated with Copper Armour™ showed a homogenous distribution of copper particles in the methacrylate matrix Fig. . AdditioStaphylococcus aureus, Pseudomonas aeruginosa, Escherichia coli O157:H7 and Listeria monocytogenes. All experiments conducted with Copper Armour™ coated samples showed a reduction, after 1 h of contact, in bacterial burden of > 99.9% compared to control samples  and the overbed Table . Additionally, the average number of cfu/100 cm2 for Staphylococcus spp. was lower on Copper Armour™ coated surfaces compared to control surfaces (Table3); however, this reduction was only statistically significant in the case of bed rails . It is important to mention that during the study S. aureus was not recovered from any surface, and only one Copper Armour™ coated surface was positive for Gram negative bacilli (720 cfu/100 cm2) and 2 for VRE (both samples with 120 cfu/100 cm2); due to these low detection rates, these microorganisms were not included in statistical analyses. In contrast, we did not observe a reduction in the average burden of yeasts / fungi on Copper Armour™ coated surfaces as compared to control surfaces. Nevertheless, the isolation of these microorganisms was sporadic overall, with values < 250 cfu/100 cm2 during the study.Copper Armour™ coated surfaces Fig.  showed a2 was lower in the case of Copper Armour™ coated surfaces compared to control surfaces  and the overbed Table   versus 33.3% (p = 0.007). Thus, Copper Armour™ exhibits antimicrobial properties able to decrease the microbial burden of high-touch surfaces in a hospital environment. Therefore, compared to control surfaces, Copper Armour™ coated surfaces were more likely to meet the threshold required for successful terminal cleaning (i.e. < 250 cfu/100 cm2), indicating that the use of this composite could contribute to schemes and practices aimed at controlling HAIs.In agreement with previous results, the frequency of samples with a microbial burden > 250 cfu/100 cmces Fig. ; this di0) Table . FurtherIt has been demonstrated that high-touch surfaces in the hospital environment are an important reservoir for infectious agents causing HAIs , 29. In Our results indicate that Copper Armour™ may be used as a self-sanitizing coating to modify existing hospital surfaces, avoiding the structural restrictions imposed by a change to solid copper. Due to its initial liquid state and subsequent hardening, this composite can be used to coat surfaces of various sizes, shapes and compositions, which reduces the cost and quantity of copper required.S. aureus, P. aeruginosa, E. coli O157:H7 and L. monocytogenes. As reported for solid copper, Copper Armour™ killed more than 99.9% of these microorganisms after one hour of contact, as well as after consecutive inoculations over 24 h . Likewise, our results showed that control bed rails had an average microbial burden of 3.323 cfu/100 cm2 (median 1.440 cfu/100 cm2) (Table 2 (median 120 cfu/100 cm2), which corresponds to a significant reduction  compared to control bed rails. Thus, Copper Armour™ exhibits antimicrobial properties able to decrease the microbial burden of high-touch surfaces in a hospital environment. Therefore, compared to control surfaces, Copper Armour™ coated surfaces were more likely to meet the threshold required for successful terminal cleaning (i.e. < 250 cfu/100 cm2), indicating that the use of this composite could contribute to schemes and practices aimed at controlling HAIs.We also evaluated the Copper Armour™ antimicrobial properties in a hospital environment. Our pilot study indicated that Copper Armour™ reduces the microbial burden of hospital surfaces, even under present day protocols of extreme hygiene. A study by Attaway et al.  showed tStaphylococcus spp. were the main bacterial group contaminating ICU bed rails. In fact, Copper Armour™ coated bed rails showed a significant  reduction in the average burden of Staphylococcus spp. compared to control bed rails  of microbial burden was observed compared to the control overbed table. Besides, a lower average burden of Staphylococcus spp. was observed in the Copper Armour™ coated overbed table compared to the control overbed table, but in this case, the reduction, while showing a trend, was not significant (p = 0.105); this is likely due to the fact that the average burden of these microorganism on the control surface was also low. Previous studies have also shown that solid copper coated overbed tables have lower level of microbial burden compared to standard surfaces [The overbed table is another Copper Armour™ coated surface in which a significant reduction  of contamination levels was observed  of control samples (Fig. It has been reported that among the objects located within a patient’s room, the IV pole shows, in general, the lowest average microbial burden , 37. Thi2. Therefore, in order to evaluate this property, it would be necessary to implement a different methodological design.The pilot study also attempted to investigate the antifungal properties of Copper Armour™. Nevertheless, we were not able to complete this aim as isolation of fungi/yeast was sporadic and with low numbers of cfu/100 cmFinally, our pilot study did not include parameters, such as whether the room was occupied / unoccupied each day or epidemiological data of the patients. Future studies, that are longer in duration and that include different hospital surfaces beyond those tested here, and that also consider patient factors are necessary to further evaluate the possible impact of Copper Armour™ on the incidence of HAIs.Our study suggests that Copper Amour TM, a novel self-sanitizing coating, exhibits bactericidal activity against important human pathogens and significantly reduces the microbial burden of hospital surfaces. Consequently, this novel composite could be used to complement infection control strategies in healthcare facilities.


To observe the risk factors affecting the occurrence of RP after gemcitabine-based induction chemotherapy.Between January 2010 and December 2017, patients with NSCLC received gemcitabine or docetaxel chemotherapy, followed by radiotherapy at Zhejiang cancer hospital were enrolled in this study. Patients were treated with gemcitabine or docetaxel induction chemotherapy, followed by radiotherapy or concurrent chemoradiotherapy. Acute radiation pneumonitis was scored post chemoradiotherapy.P = 0.005). The optimal cutoff values of lung V5, V20, V30 and MLD were set at 44% (AUC [area under the curve] = 0.593), 24% (AUC = 0.607), 14.2% (AUC = 0.622) and 1226 cGy (AUC = 0.626). On multivariate analysis, only lung V30 was identified as a predictor for grade ≥ 2 RP (P = 0.03). The grade ≥ 2 RP rate was only 9.4% for the low-risk group  in patients received gemcitabine induction chemotherapy.One hundred and eighty-four patients with NSCLC were included in the gemcitabine group and 144 in the docetaxel group. The gemcitabine group experienced a higher incidence of grade ≥ 2 RP, compared with docetaxel group (25.5% Vs. 13.2%, 30 was the best predictor of grade ≥ 2 RP.Gemcitabine chemotherapy before thoracic radiotherapy in NSCLC patients was related to a higher incidence of grade ≥ 2 RP, compared with docetaxel chemotherapy. The Lung dose-volume variable V Concurrent chemoradiotherapy is the standard treatment for locally advanced non-small cell lung cancer (NSCLC) . HistoriCurrent guidelines do not routinely recommend induction chemotherapy for patients with non-small cell lung cancer who can receive concurrent chemoradiotherapy, according to the result of CALGB 39801 . However5, V20, V30, are still the best predictors of RP at present  = 2.5) every 3 weeks for two cycles, followed by conventional fraction radiotherapy to 60 Gy combined with weekly gemcitabine (200 mg/m2). That study was closed due to excessive pulmonary toxicity. 6 of 19 patients developed grade ≥ 3 RP, and one patient died of uncontrollable RP. Furthermore, when the total dose of radiotherapy increased to 74 Gy, the incidence of fatal RP was as high as 37% [Many studies have demonstrated that gemcitabine-based chemoradiotherapy was active, with a satisfactory prognosis. However, its pulmonary toxicity was intolerable. The RP occurrence in NSCLC patients who received gemcitabine-based induction chemotherapy has been studied in several publications . In a retrospective study [P = 0.314). In our study, compared with docetaxel group, we found a higher incidence of grade ≥ 3 RP in the gemcitabine group . Furthermore, RP occurred earlier in the gemcitabine group than that in the docetaxel group (P = 0.026). The results of this study suggested that gemcitabine-based induction chemotherapy, followed by radiotherapy, was associated with a high incidence of RP. Modification of gemcitabine dosage or cycles before radiotherapy or limitation in radiotherapy planning might reduce pulmonary toxicity.Taxanes, such as paclitaxel and docetaxel, were common chemotherapeutic regimens with high efficacy in NSCLC. Comparison of taxanes based and gemcitabine bases chemotherapy has been performed in several studies. In the clinical trial CALGB 30105, patients with locally advanced NSCLC were randomly assigned to induction chemotherapy with either paclitaxel with carboplatin or gemcitabine with carboplatin followed by thoracic radiotherapy (74 Gy). The rate of grade ≥ 3 RP were 16.2 and 39.1% in the paclitaxel group and gemcitabine group, respectively. The rate of RP was significantly higher in patients who received gemcitabine induction chemotherapy than that in patients received paclitaxel  correlated significantly with RP in all patients and in the subgroup of patients who received gemcitabine induction chemotherapy. On multivariate analysis, only lung V30 was identified as an independent predictor for grade ≥ 2 RP, findings that correspond with those from several publications [30 ≥ 18% was one of the best predictors of severe acute RP among patients treated with definitive external beam radiotherapy. The observed incidence of RP was 24% for patients with V30 ≥ 18 and 6% for those with V30<18%. The cut-off value of lung V30 in our study was set as 14.2% according to the calculation the highest Youden index, findings that were slightly different from other studies. The incidence of RP was as high as 33.6% for patients with V30 ≥ 14.2 and 12.9% for patients with V30<14.2%. The cut-off values of lung V30 in patients treated with concurrent chemoradiotherapy [The lung is composed of massive parallel functional subunits. Even if some subunits were damaged by X-rays, the function of others could still operate normally. Therefore, the incidence of RP is related to the volume of the lung exceeding the lung radio-tolerant dosage. Numerous studies demonstrated that dose-volume parameters, such as lung Vications –33. The ications  showed totherapy  or adjuvotherapy  were 22 However, there were some limitations in this study. Selective bias still exists because it was a retrospective study. Secondly, radiation-induced pulmonary toxicities included radiation pneumonitis and pulmonary fibrosis. In the present study, only the occurrence and risk factors of RP were analyzed. Furthermore, because most of the patients included in this study were with lung squamous cell carcinoma, there were more male patients and fewer female patients. This limitation made the conclusions less definitive.5, V20, V30, and MLD were all associated with increased risk of grade ≥ 2 RP in patients received gemcitabine induction chemotherapy followed by radiotherapy. We also created a risk model that could be used in clinical practice. The grade ≥ 2 RP rate was only 9.4% for the low-risk group . These dosimetric parameters are recommended for NSCLC patients who received gemcitabine chemotherapy before thoracic radiotherapy.In conclusion, this study found that gemcitabine chemotherapy before thoracic radiotherapy in NSCLC patients was related to a higher incidence of grade ≥ 2 RP, compared with docetaxel chemotherapy. Lung V


Serum triiodothyronine (T3) concentration was reported to be associated with the prognosis after acute ischemic stroke. The aim of this study was to evaluate the effect of age on the prognostic value of thyroid-related hormones after an acute ischemic stroke.This was a retrospective study involving the review of 1072 ischemic stroke patients who had been consecutively admitted to the hospital within 72 h of symptom onset. Total triiodothyronine (T3), total thyroxine (T4), free T3, free T4, and thyroid-stimulating hormone (TSH) were assessed to determine their values for predicting functional outcome at the first follow-up clinic visits, which usually occurred 2 to 4 weeks after discharge from the hospital.P ≤ .001) and lower total T3 concentrations  remained independently associated with poor functional outcome in the older group. However, the independent association with poor function of lower total T3 was not confirmed in the younger group.A total of 768 patients were finally included in the study and divided into two age groups: a younger group (age < 65 years) and an older group (age ≥ 65 years). On univariate analysis, four factors—lower total T3, free T3 concentrations, higher scores on the National Institute of Health Stroke Scale (NIHSS) and the presence of atrial fibrillation—were associated with poor functional outcomes in both groups. In addition, older age, female gender, higher free T4, and lower TSH levels were also associated with poor function in the older group. On multiple logistic regression analysis, higher NIHSS scores (odds ratio [OR] =1.95; 95% confidence interval [CI], 1.66–2.30; The prognostic value of low total T3 is age-associated and more meaningful in an older population. Previous studies have shown an association of serum triiodothyronine (T3) concentration with prognosis after acute ischemic stroke; that is, a reduction in T3 level without an elevation in thyroid-stimulating hormone (TSH)  was reported to be associated with poorer clinical outcomes –4. BetteAs is well known, an individual’s levels of thyroid hormones change with age: serum total and free T3 levels are reported to decrease with age , 7, wherWe reviewed all of the available charts of patients admitted with acute ischemic stroke at the acute stroke unit of the Sir Run Run Shaw Hospital, Zhejiang University School of Medicine, between January 2011 and December 2013. Included patients met the following criteria: (1) a diagnosis of acute ischemic stroke with a new focal neurologic deficit and a corresponding lesion on magnetic resonance imaging or delayed computed tomography; on admission, non-contrast head computed tomography scan was perfomed to exclude intracranial hemorrhage; (2) admission within 72 h after symptom onset; and (3) thyroid function testing within 24 h of admission. Patients with known thyroid disease, biochemically defined overt thyroid disease, or the use of medications that affect thyroid function were excluded. To avoid any confounding effects, subjects with cancer, hematologic diseases, severe renal or liver failure, and inflammatory or infectious diseases were also excluded. All patients received standard routine treatment and intravenous thrombolysis was considered for all patients presenting within 4.5 h of symptom onset.Baseline characteristics—including demographic data; stroke risk factors, such as a history of hypertension, smoking, diabetes mellitus, hyperlipidemia, transient ischemic attacks, coronary artery disease, and atrial fibrillation; laboratory parameters routinely measured in the emergency department, including routine blood count, admission glucose, and C-reactive protein (CRP) levels; and neurologic deficits—were recorded. Blood samples for CRP were taken at the time of admission and analyzed by an immunoturbidimetric method using the Abbott Architect c16000 Biochemistry Analyzer .Thyroid function tests examined TSH, total T3, total T4, free T3, and free T4 levels. Serum concentrations of these hormones were measured using the Abbott i4000 Chemiluminescent Microparticle Immunoassay . Normal ranges of thyroid hormones are as follows: TSH, 0.35–4.94 IU/L; total T3, 0.57–1.59 ng/mL; total T4, 4.87–11.72 μg/dL; free T3, 1.71–3.71 pg/mL; and free T4, 0.7–1.48 ng/dL.The severity of stroke on admission was assessed using the National Institutes of Health Stroke Scale (NIHSS). Stroke subtype was divided into 3 groups according to the vessels involved: anterior circulation group, posterior circulation group, and both anterior and posterior circulation group. Functional outcomes were evaluated using the modified Rankin Scale (mRS) at the first follow-up clinic visit, which usually occurred 2 to 4 weeks after discharge from the hospital. Patients were classified into 2 outcome groups: poor functional outcome (mRS > 2) and good functional outcome (mRS ≤2). All of the evaluations were performed by the study neurologists.t tests and, for categorical variables, chi-square tests. Besides, spearman’s rank correlation was performed to assess the association between thyroid hormone levels and stroke severity on admission in each group. Then, the variables associated with significant differences between outcomes in each of the two groups were included in multiple logistic regression analysis and a backward likelihood ratio test was used to identify the independent predictors of poor functional outcome after stroke in each group. Receiver operating characteristics (ROC) curve analysis was used to assess discrimination and results are reported as area under the curve (AUC). All of the statistical tests were two-tailed, and P < .05 indicated statistical significance.Data were analyzed using SPSS 22.0 software for Windows. Data are expressed as medians (interquartile range [IQR]) or mean ± standard deviation (SD) for quantitative variables and as numbers (percent) for qualitative variables. Pearson correlation was used to evaluate the association of age with thyroid hormone levels. The differences in thyroid-related hormones were evaluated with the division of data into two groups based on age (< 65 years for the younger group and ≥ 65 years for the older group) , 13. Simn = 19); biochemically defined overt or subclinical thyroid disease (n = 57), defined as any of the following thyroid function tests alteration at admission: abnormal TSH, abnormal free T4 or total T4, raised free T3 or total T3 ; hematologic diseases (n = 13); severe renal or liver failure (n = 26); inflammatory or infectious diseases (n = 32); and no thyroid function test within 24 h of admission (n = 38).During the study period, 1072 ischemic stroke patients were admitted to our acute stroke unit within 72 h after symptom onset. Of these, 768 patients were included in the finally analysis and 304 patients were excluded. The reasons for exclusion were as follows: previous thyroid disease (disorder ); cancerP = .016). There was no significant difference in admission NIHSS (P = .703) and artery system involved between the two groups. Analysis of parameters related to patient history between the two groups showed that the older group included more patients with a history of previous cerebral infarction (P ≤ .001), atrial fibrillation (P ≤ .001), and hypertension (P = .001), while the younger group included more patients with a history of hyperlipidemia (P ≤ .001) and smoking (P ≤ .001). In addition, compared with the patients in the younger group, the subjects in the older group had higher levels of blood pressure on admission (P ≤ .001 for both systolic and diastolic blood pressure), with higher TSH (P = .002), free T4 (P = .011) and CRP levels (P = .001) and lower total T3 and free T3 concentrations (both P ≤ .001).The baseline characteristics of the included patients are summarized in Table r = − 0.190, P ≤ .001) and free T3  and positive correlations between age and levels of total T4  and free T4  but not TSH (P = .058).There were negative correlations between age and levels of total T3  and free T3  in both the younger and older groups. Besides, there were positive correlations between NIHSS scores and levels of total T4  in the younger group, while a negative correlation was found between NIHSS scores and TSH  in the older group.There were negative correlations between NIHSS scores and levels of total T3 (P = .282). As shown in Table At the study’s end point, a total of 38 patients (11.6%) in the younger group had poor functional outcomes with no significant difference as compared with the older group . Neither total T3 nor free T3 was independently associated with a poor functional outcome in this group. However, in the older group, in addition to the NIHSS scores , lower total T3  was also confirmed to be independently associated with poor functional outcome, even though with a less AUC [0.66 (95% CI 0.59–0.74) than NIHSS [0.87(95% CI 0.85–0.88)]. This finding indicates that the prognostic value of a lower total T3 is age-associated and more meaningful in an older population.On multiple logistic regression analysis, only the NIHSS scores remained independently associated with poor functional outcome in the younger group (OR = 1.56; 95% CI 1.39–1.76; This study demonstrates that in euthyroid patients with acute ischemic stroke, the association of total T3 level with functional outcome differs with age. In patients at or above 65 years of age, a lower total T3 concentration was an independent predictor of poor functional outcome after ischemic stroke. However, none of the thyroid–related hormones, including total T3, were independently associated with a poor functional outcome in patients below 65 years of age.Low T3 syndrome and lower T3 serum concentrations are common complications in acute cerebrovascular disease  and are T3 has been shown to have neuroprotective effects in animal models. Several attempts have been made to directly investigate the neuroprotective actions of T3 in mice models of transient middle cerebral artery occlusion –21. In tOn the other hand, it is well known that, within a few hours after the onset of stroke, plasma T3 decreases, and the magnitude of the change is related to the stroke severity –25. The Ischemic stroke increasing in incidence with age and tends to present with more severe symptoms; it also has worse outcomes in the elderly compared with younger patients . As a spSeveral limitations should be considered in the interpretation of our study. First, this is a retrospective study, which poses the risk of selection bias. Second, we lacked long-term clinical follow-up data on our patients. Third, we only use a mRS cut-off value to evaluate the functional outcome after stroke. One previous study has shown that reduced T3 concentrations can serve as an independent biomarker of worse cognitive outcomes after ischemic stroke, a common complication in stroke patients . FurtherThis study assessed the effect of age on the prognostic value of thyroid-related hormones in patients with acute ischemic stroke. We found that the prognostic value of lower total T3 for poor outcome after ischemic stroke may not always be confirmed in all age populations and that, instead, it may be more clinically meaningful in older patients. Therefore, selecting a population of the appropriate age in order to study the prognostic value of candidate predictors is reasonable.