Paeonia lactiflora root (baishao in Chinese) is a commonly used herb in traditional Chinese medicine (TCM). Paeoniflorin (PF) and albiflorin (AF) are two major active constituents of P. lactiflora. In this paper, we aimed to investigate the hematopoietic effects of PF and AF on myelosuppression mice induced by radiotherapy and to explore the underlying mechanism. The finding indicated that PF and AF significantly increased the numbers of white blood cells (WBC) and reversed the atrophy of thymus. Furthermore, PF and AF increased the levels of granulocyte-macrophage colony-stimulating factor (GM-CSF) and interleukin-3 (IL-3) and reduced the levels of tumor necrosis factor-α (TNF-α) in serum and increased the level of colony-stimulating factor (G-CSF) in plasma. Lastly, PF and AF not only enhanced the mRNA levels of GM-CSF and G-CSF in the spleens, but also increased the protein levels of G-CSF and GM-CSF in bone marrow. Our results suggest that PF and AF may promote the recovery of bone marrow hemopoietic function in a myelosuppressed mouse model. Paeonia lactiflora Pall (family Ranunculaceae) is commonly used in China, Korea, and Japan. P. lactiflora is a component herb of many traditional formulae, such as Siwu-Tang, which has been widely used in treating palpitation, dysmenorrhea, chronic inflammation, and aminia [ P. lactiflora that have been identified, including paeoniflorin (PF), albiflorin (AF), oxypaeoniflorin, paeonilactinone, benzoyloxypaeoniflorin, and lactinolide [ P. lactiflora containing paeoniflorin and albiflorin (CPA) showed ameliorative effects on myelosuppression induced by radio and chemotherapy [The root ofd aminia . There atinolide . It is wtinolide , 4, musctinolide , 6, infltinolide , 8, and tinolide , 10. Howtinolide –13. In aotherapy . In anotα (TNF-α), that can be used to accelerate hemopoietic recovery.Blood/bone marrow system is one of the largest organs in the body that is an important potential target in ionizing radiation . Acute e P. lactiflora, have the hematopoietic effect as well as the antimyelosuppression effect. Mechanisms associated with hematopoietic cytokines may also be involved in the hematopoietic effects of the two isomers. To test this hypothesis, we examined the effects of PF and AF in 3.5 Gy Co60  γ-rays irradiated-induced myelosuppression mice [Consistent with our previous study –13, stilion mice . We alsoion mice , the thyPaeoniflorin (PF) and albiflorin (AF) were prepared in our library . The purity of PF (purity = 98.6%) and AF (purity = 96.7%) was measured by reverse-phase high-performance liquid chromatography (HPLC) coupled with ultraviolet detection according to Chinese Pharmacopoeia . The HPLC chromatogram and chemical structures of the two isomers are shown in Kunming mice  were obtained from the Vital River Co., Ltd.  and maintained at controlled temperature (22 ± 2°C) and humidity (50 ± 10%) with a 12 h light/dark cycle. All efforts were made to minimize the pain of animals. The experimental protocol was approved by the Ethics Committee of Beijing University of Chinese Medicine (number Kj-dw-18-20150731-01).γ-ray treated myelosuppression mice model was set up according to Liang's method [n = 10 mice per group, males), including (1) normal control group (control), (2) γ-ray treated group (model), (3) high-dose PF treatment group , (4) L-dose PF treatment group , (5) high-dose AF treatment group , and (6) L-dose AF treatment group .This s method , 18. Aft60  γ-rays at a dose rate of 1.60 Gy. After irradiation, control group mice were treated in exactly the same way as the irradiated animals; PF and AF were administered daily intragastrically at a dose of 0.2 mL/20 g body weight for 14 consecutive days. Normal and model mice were given normal saline.All mice except the control and model were intragastrically (i.g.) pretreated with the different doses of chemical compounds, PF (30 mg/kg and 15 mg/kg), and AF (30 mg/kg and 15 mg/kg) for 7 days. On the 8th day, mice except the normal group received total body irradiation of 3.5 Gy CoμL) of blood with EDTA (50 μL) was used for peripheral hemogram analysis by Beckman Coulter Ac and T 5 full-automatic blood cell analyzer . After the mice were sacrificed, the thymus gland and spleen were excised and weighted to calculate the thymus index and spleen index: (organ weight/body weight) × 1000.After the last administration, body weights of mice were measured. Then the mice were subjected to light diethyl ether anesthesia and blood was collected into clean test tubes with or without ethylenediaminetetraacetic acid (EDTA) by extracting eyeballs. A portion  kits .Plasma or serum samples were collected from the sacrificed mice, and the levels of G-CSF, GM-CSF, IL-3, IL-6, and TNF-Ct method and calculated setting normal control as 1.The spleen tissues were homogenized and the total RNA was extracted from the supernatant fraction. Then total RNA from each sample was reverse-transcribed into cDNA using a Super RT cDNA kit , and the synthesized cDNA was used for RT-qPCR amplification using SYBR green Real-time PCR Master Mix. Furthermore, the nucleotide sequences of forward and reverse primers used for PCR are shown in α .Three mice in each group were sacrificed and then bone marrow from the femur was collected. The total protein in bone marrow was extracted with lysis buffer , and 50 mg protein was resolved on a 10% sodium dodecyl sulfate polyacrylamide gel. The fractionated proteins were electrophoretically transferred to an immobilon polyvinylidene difluoride membrane and probed with antibody of G-CSF, GM-CSF, IL-3, IL-6, and TNF-t-tests. A P value < 0.05 indicates a statistically significant difference.Results are expressed as mean ± SD. Statistical significant differences were determined by one-way analyses of variance and Student's P < 0.001). PF-H or AF-H treatment significantly elevated the number of WBC (P < 0.001). AF-L group also increased the number of WBC significantly (P < 0.01).As shown in P < 0.01, P < 0.001). PF-H significantly increased the body weight (P < 0.01) and thymus index (P < 0.05) and AF-H also significantly increased the body weight (P < 0.01) and thymus index (P < 0.05). AF-L group increased the body weight (P < 0.05). It proved that both PF and AF could reverse the loss of body weight and the atrophy of hemopoietic organ  induced by irradiation. There is no change on spleen index.As shown in As shown in P < 0.001, P < 0.001), while levels of GM-CSF and G-CSF in PF-H  and AF-H significantly increased , respectively. However, PF-L significantly increased G-CSF (P < 0.01) and AF-L increased GM-CSF (P < 0.01). Irradiation induced a decrease of IL-3 in serum (P < 0.001), and PF-H and AF-H administration increased the level of IL-3 compared to the model . As shown, irradiation also induced an increase of TNF-α compared to control (P < 0.01), and PF-H and AF-H administration decreased the level of TNF-α, respectively .As shown in P < 0.01) in the model animals. Compared to the model group, the mRNA level of G-CSF was elevated in PF-H (P < 0.01) and in AF-H (P < 0.001) treatment groups. Besides, there were no significant differences between the normal and the model mice on the mRNA levels of GM-CSF, IL-3, IL-6, and TNF-α. PF-H increased the mRNA levels of GM-CSF and TNF-α . AF-H significantly increased the mRNA levels of GM-CSF and IL-3 . The data suggested that PF-H and AF-H could significantly enhance the mRNA levels of GM-CSF, G-CSF, and TNF-α in the spleens.As shown in P < 0.01, P < 0.001, and P < 0.01) and AF-H also could increase the levels of G-CSF, GM-CSF, and IL-3 compared to the control group, respectively . As shown, irradiation induced increases of IL-6 and TNF-α after irradiation compared to the sham irradiation , PF-H could increase the levels of IL-6 and TNF-α compared to model group , and AF-H significantly increased IL-6 and TNF-α levels compared to control group . Interestingly, the IL-6 and TNF-α protein levels were opposite to those in serum.As shown in Mice exposure to ionizing radiation leads to injury to the lymphoid and haemopoietic system, which can result in septicaemia and death . Myelosuα. A key feature of enhancing recovery from the radiation syndrome is the hematopoietic cytokines activation [α is a potent inhibitor, which in turn affects the differentiation of early bone marrow progenitor cells by altering their response to CSFs [α were opposite to those in serum, and the reason needs to be investigated further. However, IL-6 is produced by a number of normal and transformed cell lines [ Panax ginseng, which is traditionally used as a restorative, antidiabetic, antivomiting, and anticancer agent worldwide) or Fu fang E'jiao Jiang (Panax ginseng is a component herb of this formula) changed the expression of hematopoiesis-related cytokines in animal models [In addition, hematopoiesis is a diverse process, regulated by various hematopoietic cytokines, such as G-CSF, GM-CSF, IL-3, IL-6, and TNF-tivation . In thistivation . We alsotivation  and accetivation . Further to CSFs  and acce to CSFs . Interesll lines , which cll lines . Howeverl models , 27. PF In conclusion, the results of the present study indicate that PF and AF have potent ameliorative effects on radiation-induced myelosuppression mice. PF and AF stimulate the hematopoiesis-related cytokines and enhanced hematopoietic progenitor cell activity resulting in accelerated blood cell recovery. Additionally, further pharmacological studies to determine more mechanisms underlying the therapeutic effects of PF and AF are needed.


Many women wear facial makeup to accentuate their appeal and attractiveness. Makeup may vary from natural (light) to glamorous (heavy), depending of the context of interpersonal situations, an emphasis on femininity, and current societal makeup trends. This study examined how light makeup and heavy makeup influenced attractiveness ratings and facial recognition. In a rating task, 38 Japanese women assigned attractiveness ratings to 36 Japanese female faces with no makeup, light makeup, and heavy makeup (12 each). In a subsequent recognition task, the participants were presented with 36 old and 36 new faces. Results indicated that attractiveness was rated highest for the light makeup faces and lowest for the no makeup faces. In contrast, recognition performance was higher for the no makeup and light make up faces than for the heavy makeup faces. Faces with heavy makeup produced a higher rate of false recognition than did other faces, possibly because heavy makeup creates an impression of the style of makeup itself, rather than the individual wearing the makeup. The present study suggests that light makeup is preferable to heavy makeup in that light makeup does not interfere with individual recognition and gives beholders positive impressions. Many women wear facial makeup in their daily lives to accentuate their attractiveness and create favorable impressions. However, what type of makeup currently creates the most appealing and memorable impressions is not always evident. If people were aware of the social and psychological effects of makeup on the beholder, they would be more satisfied with, and knowledgeable about, their selection and use of cosmetic products.The positive bias toward attractive individuals is known as the beauty halo effect or a beauty-is-good stereotype . For exaHowever, the type of makeup that is considered most attractive to others is an unsettled issue. People’s positive attitudes toward the attractiveness of another’s makeup are affected by various factors, including societal and interpersonal contexts. For example, light makeup that enhances natural facial expressions is often considered attractive or appropriate in daily situations, whereas glamorous heavy makeup, which has greater contrasts as well as obvious coordination of the makeup applied to the eyes or lips with clothing or hairstyle, is considered more appropriate at events such as parties or celebrations. A previous study suggests that color cosmetics can be considered phenotypic extensions that are linked to biological meanings . Moreovethe Act on Securing, Etc. of Equal Opportunity and Treatment between Men and Women in Employment in 1986, many working women have assumed roles in public life that are comparable to those of business men. The makeup trends at that time leaned toward heavy eyebrows and strongly accented facial features. Thus, working women during that period tended to wear heavy makeup. More recent trends in makeup have appeared since the Great East Japan Earthquake in 2011; these reveal a shift toward natural, lighter makeup. One interpretation of this trend is that lighter makeup expresses femininity or beauty in the search for healing during a time of confusion. Specific features of this trend can be seen in the increased brightness of the preferred colors for eyebrow accents and lip gloss; moreover, there is a growing emphasis on the expression of one’s personality as an element of natural femininity  retention period. Here we define the light makeup is a kind of makeup that is characterized by naturalness and femininity. Reddish colors were used and blended naturally into the skin. On the other hand, we define the heavy makeup as a kind of makeup that is characterized by perfectness, maturity, and coolness. Dark, low-chromatic colors were used to enhance the luminance contrast in the face. A total of 40 adult women were recruited by a research firm and paid for participation. The conditions of recruiting were (1) physically and mentally unimpaired, (2) being right-handed, (3) having reported normal or corrected to normal vision, (4) wearing makeup on a daily basis, and (5) being familiar with the effect of makeup on the impression of a face. Participants were gave written informed consent before testing. The Ethical Committee of Shiseido Global Innovation Center approved this study. Due to a technical failure in the experiment, the data of two participants could not be used. The following analysis was conducted on the data of 38 women (mean age: 27.1 ± 4.8 years).Figure 1 shows examples of facial images. The quality of all the pictures was checked carefully by a professional makeup artist and a makeup product manager. When necessary, the pictures (including no makeup faces) were retouched by computer graphics to meet a standard of cosmetic quality and naturalness. The size of each picture was 500 × 368 pixels. A constant-size oval occluding window was applied to each photograph to hide the background and hair. The 72 facial images were divided in two sets of 36 faces (12 for each type).A total of 72 female face images with neutral facial expressions were used. Among them, 68 faces were of different female models  and four faces were generated by using computer graphics. Even when these virtual faces were excluded from the analysis, the conclusion did not change. Therefore, the results of the whole dataset are reported here. There were three categories of faces: no makeup, light makeup, and heavy makeup. These faces were of different individuals. For the light makeup, foundation was blended smoothly into the skin tone. The outline of the facial features was blurred and the shape of the eyebrows was smoothed to create softness. The eye shadow was also blurred to blend into the skin tone. The lips were tinged with a reddish gloss. A soft red was used on the cheeks and blended from the center into the skin at the periphery. For the heavy makeup, a matte foundation covered the skin. The outline of the facial features was emphasized with straight lines. The eyebrows were drawn in a dark color with sharp, straight lines to make the center of the brow higher. Dark eye shadow was applied to produce long, slitted eyes. A matte brownish color with a sharp outline was added to the lips. Cheek color was a dark red–brown and blurred so as to accent the cheekbone. The stimuli were presented on a computer screen  in a dimly lit room by Inquisit 4.0 (Millisecond software). All the faces were displayed against a black background and subtended 8° wide × 10° height at a viewing distance of 70 cm.Figure 2 shows a schematic diagram of the experiment. The experiment consisted of two tasks. In a rating task, 36 images consisting of 12 no makeup, 12 light makeup, and 12 heavy makeup faces were presented one at a time. Each image appeared only once. After viewing a frame for 1500 ms, a face was displayed for 1000 ms. Subsequently, a blank frame appeared for 1000 ms, and then the rating screen was presented. The participants were instructed to rate each face for attractiveness by moving and clicking a computer mouse using a 7-point scale as follows: 1 = extremely unattractive, 2 = very unattractive, 3 = moderately unattractive, 4 = neither, 5 = moderately attractive, 6 = very attractive, 7 = extremely attractive. The next trial began 1000 ms after the response. The stimuli were presented in randomized order across participants.not at all confident, 2 = somewhat confident, and 3 = extremely confident) by button press. The next trial began 2000 ms after the confidence rating. The response time was not limited in either task. Before each task, the participants performed a practice trial with male facial images to ensure they were able to perform the task.The recognition task started with an interval of 3 minutes after the end of the rating phase. No advance notice was given for the recognition task. All the 72 images were presented one by one in randomized order; half were previously viewed images (“old”) and the other half were unviewed images (“new”). After viewing a frame for 1500 ms, a face was displayed for 1000 ms. The participants were asked to indicate whether they recognized the face image by choosing either the “old” or “new” button on a response pad (Cedrus RB-530). The response hands were counterbalanced across participants. After a button press, a confidence rating screen appeared on which participants rated their recognition confidence using a 3-point scale  and response bias [Br = False recognition/(1 – Pr)] were also calculated  rates, correct rejection rates, and confidence ratings were analyzed. In addition, the geometric means of correct response times  were calculated. A receiver operating characteristic (ROC) curve was plotted for each makeup category and each participant. Each curve consisted of five points, which were calculated from five response criteria determined by a combination of the yes/no answers and confidence scores. Specifically, a “yes” answer with a high confidence score of three was regarded as the “yes” response at the highest response criterion, and a “yes” answer with a high or middle confidence score was regarded as the “yes” response at the second highest response criterion, and so on. Correct and false recognition rates were calculated for these five response criteria. The area under the ROC curve (AUC) was calculated as a measure of the sensitivity of face recognition. According to the previous study, the measures of discrimination accuracy  = 17.83, p < 0.001, Pr) was lowest for heavy makeup faces. Again, a linear effect across conditions was significant, F = 8.18, p = 0.007, Br) differed significantly across the categories, F = 17.63, p < 0.001, Post hoc pair-wise comparisons showed that Br was higher for heavy makeup faces than for no makeup and light makeup faces, ps < 0.05. Moreover, heavy makeup faces were different from the other faces in terms of the reaction time and confidence rating data. Heavy makeup faces were associated with the shortest correct recognition (hit) time and the longest correct rejection time. Correct recognition time and correct rejection time differed significantly across the categories, F = 7.57, p = 0.002, F = 4.24, p = 0.022, Post hoc comparisons showed that correct recognition was faster for heavy makeup than for light makeup and that correct rejection was faster for heavy makeup than for no makeup, ps < 0.05. Confidence in correct rejection also differed significantly across the categories, F = 4.44, p = 0.019, Post hoc comparisons showed that heavy makeup faces were judged with lower confidence as compared to light make up faces, p < 0.05.For the recognition task, hit and correct rejection rates differed significantly between the categories: the hit rate was higher and the correct rejection rate was lower for heavy makeup than for no makeup and light makeup. In the impression reporting session at the end of the experiment, both light and heavy makeup faces were chosen more frequently than no makeup faces as faces that made an impression. The rate of choice was 27.4% for no makeup, 34.7% for light makeup, and 37.9% for heavy makeup. The reasons for choosing these faces differed across makeup categories: asymmetry, close-set eyes, acne, and moles for no makeup faces; beautiful, gentle-looking, and feminine for light makeup faces; and thin, haughty, and unfavorable for heavy makeup faces.The present study examined the effects of light and heavy makeup on the rating of attractiveness and face recognition. The results showed that facial attractiveness was judged greatest for faces with light makeup. Heavy makeup was in second place, and no makeup received the lowest ratings. The accuracy of facial recognition was higher for faces without makeup than for faces with light or heavy makeup. Therefore, facial distinctiveness was shown to be more associated with facial recognition than with facial attractiveness. In addition, the response bias for heavy makeup faces was significantly more liberal compared with no makeup and light makeup faces. That is, faces with heavy makeup were often recognized falsely. In the impression reporting session at the end of the experiment, the participants replied that the heavy makeup stood out in their memory and it made as much of an impression as light makeup. Nevertheless, the results showed that facial recognition was in fact lower for the heavily made-up faces than for the other faces.Attractiveness was rated higher for faces with makeup than for faces without makeup. Makeup, whether light or heavy, creates a positive impression on observers by concealing and minimalizing the negative elements of one’s natural face. This is often a primary reason why women wear makeup on a daily basis. For instance, facial features that are commonly considered negative  include asymmetric features, the shape of one’s eyes or lips, and uneven skin tone. The texture of facial skin affects the perception of a subject’s sex . Aging hIn this study, faces with light makeup were judged as more attractive than those with heavy makeup. In a previous study , faces wThe accuracy of face recognition was higher for faces with no makeup and light makeup than for faces with heavy makeup. Natural faces have idiosyncratic features. For example, Interestingly, poor memory performance for heavy makeup faces was associated with a liberal response bias. Novel faces with heavy makeup were often recognized falsely as if they were faces the observer had previously seen. This is possibly because such makeup obscures individual facial features, and the distinctiveness of the makeup itself becomes prominent. That is, participants had a strong memory for the style of makeup but they were not able to identify individual faces. This effect of heavy makeup may lead to an interesting speculation. In the past, working women in Japan often wore heavy makeup. Before the enactment of laws protecting women in the workplace, women were not a major presence in the workforce. Heavy makeup may have been an effort to increase the recognition of women in work places. In a sense, it was effective. However, at the same time, heavy makeup has an untoward effect: it obscures individual characteristics. Currently, women have essential roles in many work places. Therefore, light makeup may become more popular and more effective because it facilitates individual recognition. Of course, depending on circumstances and purpose, heavy makeup may be more effective as an element of interpersonal communication than light makeup; in some situations, heavier makeup may even be considered more attractive.Applying facial cosmetics affects women’s self-images positively . The resThere are several limitations in this study. First, the faces featuring no makeup, light makeup, and heavy makeup were of different individuals. Therefore, the distinctiveness of individual faces might affect recognition performance. It is very costly to prepare three types of makeup for each individual model while maintaining a high standard of the quality of photographs. However, the effect of makeup on a single individual face needs to be tested in a future study. Second, facial makeup images used in the present study were digitally retouched. Although the quality of the final images was carefully controlled by a professional makeup artist, actual makeup and digital manipulation might have different effects. This issue will be resolved by replicating this study using models with actual makeup alone. Third, only female participants were recruited in this study. In connection to the behavior of partner selection, it is worth examining whether men show a similar pattern of results.The present study assessed the effects of light and heavy makeup in determining perceived facial attractiveness and face recognition using an old/new task. The results showed that the highest ratings of attractiveness were for faces with light makeup; in addition, recognition accuracy was highest for faces with no makeup and lowest for faces with heavy makeup. These results show that the distinctiveness of a face, rather than its attractiveness, has a greater effect on recognition accuracy. Moreover, it was found that people tended to mistake unseen faces with heavy makeup for previously presented ones. The present study suggests that, at least under current fashion trends, light makeup is preferable to heavy makeup because it is viewed as more attractive and allows for greater expression of individual personality and easier recognition by others.KT and HN planned, collected, analyzed, interpreted the data, and wrote the paper. HO contributed for the preparation of stimulus.KT and HO are employed by Shiseido Global Innovation Center, Shiseido Co., Ltd., a company which manufactures cosmetics, and HN serves as a technical advisor for Shiseido Co., Ltd. There are no patents, products in development or marketed products to declare.


Myelin oligodendrocyte glycoprotein antibody (MOG Ab) associated demyelination represents a subgroup of autoimmune demyelination that is separate from multiple sclerosis and aquaporin 4 IgG-positive NMO, and can have a relapsing course. Unlike NMO and MS, there is a paucity of literature on immunopathology and CSF cytokine/chemokines in MOG Ab associated demyelination.To study the differences in immunopathogenesis based on cytokine/chemokine profile in MOG Ab-positive (POS) and -negative (NEG) groups.We measured 34 cytokines/chemokines using multiplex immunoassay in CSF collected from paediatric patients with serum MOG Ab POS  and serum MOG Ab NEG  demyelination. We generated normative data using CSF from 20 non-inflammatory neurological controls.The CSF cytokine and chemokine levels were higher in both MOG Ab POS and MOG Ab NEG demyelination groups compared to controls. The CSF in MOG Ab POS patients showed predominant elevation of B cell related cytokines/chemokines  as well as some of Th17 related cytokines (IL-6 AND G-CSF) compared to MOG Ab NEG group . In addition, patients with elevated CSF MOG antibodies had higher CSF CXCL13, CXCL12, CCL19, IL-17A and G-CSF than patients without CSF MOG antibodies.Our findings suggest that MOG Ab POS patients have a more pronounced CNS inflammatory response with elevation of predominant humoral associated cytokines/chemokines, as well as some Th 17 and neutrophil related cytokines/chemokines suggesting a differential inflammatory pathogenesis associated with MOG antibody seropositivity. This cytokine/chemokine profiling provides new insight into disease pathogenesis, and improves our ability to monitor inflammation and response to treatment. In addition, some of these molecules may represent potential immunomodulatory targets. Myelin oligodendrocyte glycoprotein (MOG) is a myelin antigen located at the outer surface of the central nervous system (CNS) myelin sheath, and is a target for autoimmune responses that results in CNS inflammation and demyelination . Using cIn vitro experiments have shown that patient serum containing MOG antibodies can induce complement, natural killer cell and antibody dependent cell mediated toxicity, and disrupt oligodendrocyte cytoskeleton  and 9 patients with MOG Ab NEG (ADEM = 5 and TM = 4) demyelination were included. We ensured that MOG Ab POS and NEG groups were clinically similar with regard to proportions of ADEM and TM. Patients with ADEM and TM fulfilled International Pediatric MS Study Group (IPMSSG) diagnostic criteria 2013 . Twenty The radiological abnormalities on acute and follow up MRI brain including extent and configuration of lesions were recorded and rated by neuroradiologist (KP) blinded to diagnosis according to the previously published protocol by Baumann et al . The seq® MAP system, Millipore Corporation, Missouri U.S.A.) according to the manufacturer’s instructions, using a combination of 23-plex (MPHCYTOMAG60K23), 6 plex (MPHCYP2MAG62K06), and 3-plex (MPHCYP3MAG63K03) Millipore Human Cytokine panel kits. The 23-plex kit contained antibody-conjugated beads for following cytokines and chemokines: IL-1ra, GM-CSF, IL-1b, TNF-α, IL-2, IL-4, IL-6, IL-8, IL-10, IL-13, IL-17A, IFN-γ, CCL2/MCP-1, CCL5/RANTES, CXCL1/GRO, CXCL10/IP-10, CCL3/MIP-1a, CCL4/MIP-1b, IL-12 (p40) and IL-12 (p70), IFN-α, G-CSF and CCL11/Eotaxin. The 6-plex kit was used to detect IL-21, IL-23, CXCL13/BCA-1, CCL17/TARC, CCL21/6Ckine and CXCL12/SDF-1. The 3 Plex kit contained antibody-conjugated beads for CXCL9/MIG, CXCL11/I-TAC, and CCL19/MIP-3b. For each assay, the curve was derived from various concentrations of the cytokine standards assayed in the same manner as patient samples. All samples were measured undiluted. The methodological details including assay method, lower detection limits and coefficient of variance are available at the manufacturer’s website, “www.merckmillipore.com”Thirty-two cytokines were measured by multiplexed fluorescent bead-based immunoassay detection (MILLIPLEXAPRIL (sensitivity <2pg/ml) and BAFF (sensitivity 0.4ng/ml) were performed using ELISA kits as per the manufacturer’s instructions . All samples were analyzed undiluted except 6 samples, where volume was insufficient for BAFF analysis. In these cases, results were multiplied by the appropriate dilution factor.Statistical analysis was performed using R statistical programme and graphs were composed using Graph Pad Prism software version 6. We compared clinical and demographic features between MOG Ab POS and NEG groups using Mann-Whitney test. The CSF cytokine/chemokine data were analyzed to check for a normal distribution. As data did not show Gaussian distribution, statistical analyses of cytokine/chemokine levels were performed using Kruskal-Wallis test for multiple groups and Mann-Whitney test for two groups and 2 tailed p value was calculated. No adjustment was made for multiple group statistical comparisons, as this study was largely exploratory with an intention to study the immunopathological mechanisms based on CSF cytokines/chemokines. The p-values from these analyses should be viewed as a measure of the strength of evidence for each association. We examined the correlations between cytokine/chemokine levels in the CSF and with disability score during admission and follow up by the Spearman rank correlation test. In the correlation analysis, p values <0.05 were considered to indicate statistical significance.6 /L), CSF neutrophils (median 5 vs 1 106 /L), CSF IgG (median 0.04 vs 0.025 g/L), and protein (median 0.42 vs 0.29 mg/dl) levels were higher in the MOG Ab POS CSF samples than in the MOG Ab NEG (The patients in MOG Ab POS and control group were younger compared to MOG Ab NEG group (median 4.7 vs 4.9 vs 9.3 years). The time to CSF sampling from onset of neurological symptoms was shorter in the MOG Ab NEG group compared to MOG Ab POS group (median 4 vs 6 days). The ESR was more commonly elevated in MOG Ab POS group. The CSF cell count , larger white matter lesion size, contrast enhancement of lesions, T1 hypointense lesions, optic nerve involvement, and more residual signal changes at follow up (FU) MRI compared to MOG Ab NEG group  were elevated in the serum MOG Ab POS group compared to MOG Ab NEG group.All five of the five measured CSF B cell associated cytokines/chemokine levels were elevated in serum MOG Ab POS group compared to MOG Ab NEG group .CSF IL-10 was elevated in MOG Ab POS group compared to MOG Ab NEG group.Of the 13 other CSF cytokines/chemokines measured, only IL-1ra levels were higher in serum MOG Ab POS group compared to MOG Ab NEG group.The CSF cytokines/chemokines which were elevated (p<0.05) in serum MOG Ab POS compared to MOG Ab NEG are presented in Nine of the 10 serum MOG Ab POS patients had adequate CSF available for CSF MOG antibody testing. Four of these 9 samples were positive for CSF MOG Ab, and 5 were CSF MOG Ab negative. All 34 CSF cytokines and chemokine levels were compared, of which only median concentrations of B cell  and Th17  cytokines were higher (p <0.05) in CSF MOG Ab POS group compared to CSF MOG Ab NEG group .There were no correlations of cytokines/chemokines with disability outcome using modified Rankin scale at admission and follow up. Most cytokine/chemokine levels decreased with longer duration of CSF sampling after onset of neurological symptoms. The majority of B cell cyto/chemokines correlated with each other and with Th17 molecules  and IL-10, but did not show any relation with Th1 (except TNF-α) and Th2 related cytokines/chemokines . As previously reported, the serum MOG Ab POS patients in our cohort were younger at presentation, had higher systemic inflammatory markers (ESR), higher CSF cell count, CSF protein, CSF IgG levels, higher lesion load on MRI and more often had relapses and minor neurological deficits at follow-up which suggests that MOG Ab associated demyelination is a more inflammatory CNS syndrome than MOG Ab negative demyelination , 12, 25.in vitro studies [We investigated a comprehensive array of cytokines/chemokines in our patients with seropositive and seronegative MOG Ab associated demyelination. Although Th1 and Th2 cytokines were often elevated in both MOG Ab POS and MOG Ab NEG groups compared to controls, there was no significant difference between the serum MOG Ab POS and MOG Ab NEG groups except for TNF-α. Interestingly, TNF-α has been shown to be produced by both Th1 and Th17 cells in  studies .All of the measured CSF B cell associated chemo/cytokines  were higher in the serum MOG Ab POS group compared to MOG Ab NEG group. These chemo/cytokines play an important role in recruitment, clonal selection and expansion of B cells –31. In ain vivo regulatory response to counteract proinflammatory cytokines [Th17 related cytokines/chemokines, predominantly IL-6 and G-CSF, were significantly elevated in serum MOG Ab POS group compared to MOG Ab NEG group. Recently IL-6 was reported to correlate with MOG antibody titers in paediatric patients with monophasic acquired demyelination syndromes . IL-6 acytokines .in vivo . And finally, we could not perform comparisons between serum and CSF samples to determine whether the cytokine/chemokines are predominantly produced intrathecally or peripherally, due to the lack of available stored frozen serum samples.In summary, our study points to a role of predominant humoral immunity and possible role of Th17 cells and neutrophils in MOG antibody associated demyelination, which strengthens the hypothesis that MOG antibody defines a separate immunopathological syndrome with dominant humoral autoimmunity.S1 FileTable A. Details of clinical syndrome, treatment and outcome of MOG Ab POS and NEG patients with acute demyelination episode. Table B. Comparison of radiological abnormalities between MOG Ab POS and MOG Ab NEG demyelination groups.(DOCX)Click here for additional data file.


Here, we present clinical follow-up of all patients (n=62) who participated in these trials.Currently, there is no adjuvant treatment available that has been shown to reduce the chances of recurrence after surgical excision of localized melanoma. Between 2002 and 2007 we conducted three randomized and placebo  controlled Phase II clinical trials in which we treated clinically stage I-II melanoma patients with 1) GM-CSF 3 μg/kg), 2) CpG-B , and 3) CpG-B, alone or combined with GM-CSF , through 1-4 intradermal injections, directly adjacent to the primary melanoma excision scar, within a week of re-excision and sentinel lymph node (SLN) biopsy[ μg/kg, 2Upon pathological examination of the SLN, a remarkable difference in numbers of pathologically stage III patients was revealed, with significantly less tumor positive SLN in the CpG-B/GM-CSF administered groups , suggesting a rapidly induced antitumor response. Follow-up further provided evidence of systemic tumor protection: in the CpG/GM-CSF-treated groups only two recurrences occurred, both within 26 months of SLN biopsy, whereas nine recurrences were observed in the saline group (range 5-113 months). In post-hoc analysis, at a median follow-up of 79 months, this translated into a significantly longer recurrence-free survival (RFS) in the CpG-treated group (p=0.010). Even for patients with pathologically confirmed and prognostically favorable stage I-II, this difference held true (p=0.017). Disease-specific survival rates at this time were 76.9% in the saline vs. 94.5% in the CpG-B/GM-CSF group.We conclude that intradermally administered CpG-B/GM-CSF is safe and may prolong RFS. Consistent with findings from immune monitoring, apparent down-staging and prolonged RFS may result from the respective boosting of local and systemic antitumor immunity through local conditioning of the SLN. These exciting findings warrant further clinical exploration of local non-toxic immune potentiation in early-stage melanoma to prevent disease recurrence and metastatic spread.


There is a number of research work in the literature that have applied sEMG biofeedback as an instrument for muscle rehabilitation. Therefore, sEMG is a good tool for this research work and is used to record the myoelectric activity in the paraspinal muscles of those with AIS during habitual standing and sitting. After the sEMG evaluation, the root-mean-square (RMS) sEMG values of the paraspinal muscles in the habitual postures reflect the spinal curvature situation of the PUMC Type Ia and IIc subjects. Both groups have a stronger average RMS sEMG value on the convex side of the affected muscle regions. Correction to posture as instructed by the physiotherapist has helped the subjects to achieve a more balanced RMS sEMG ratio in the trapezius and latissimus dorsi regions; the erector spinae in the thoracic region and/or erector spinae in the lumbar region. It is, therefore, considered that with regular practice of the suggested positions, those with AIS can use motor learning to achieve a more balanced posture. Consequently, the findings can be used in less intrusive early orthotic intervention and provision of care to those with AIS. Adolescent idiopathic scoliosis (AIS) is a multifactorial, three-dimensional deformity of the spine and trunk. It can appear and sometimes progress during any of the rapid periods of growth in children . NoninvaBiofeedback is a nonmedical process that involves the measuring of specific and quantifiable bodily functions of a subject, such as the brain wave activity, blood pressure, heart rate, skin temperature, sweat gland activity, and muscle tension, thus conveying the information to the patient in real-time. The basic aim of biofeedback therapy is to support a patient in realizing his/her self-ability to control specific psychophysiological processes . The litDifferent studies have also been done on the paraspinal muscle activity of adolescents with idiopathic scoliosis by using sEMG. Avikainen et al. recorded the force-time and the EMG-time curves of the paraspinal muscles during maximal isometric trunk extensions. The isometric force-time curves as well as the maximal integrated EMG activity that were recorded from both sides of the thoracic and lumbar spines did not show any significant differences between the normal and scoliotic groups . Chwała A screening program was carried out in Hong Kong during 2014 with 2 schools and the target population was 10–13-year-old females. During the examination process, the subjects were invited to perform Adam's forward bending test and an OSI scoliometer was employed to measure the rib hump which is directly related to spinal rotation and rib deviations. The angle of trunk rotation (ATR) in the spine of the subjects was measured while lying prone in order to preliminarily assess their spinal conditions  were divided into 3 groups: PUMC type Ia, a single thoracic curve with apex between the T2 and T11-T12 disc; PUMC type Ib, a single thoracolumbar curve with apex at T12, T12-L1 disc, and L1; and PUMC type IIc (double curves) thoracic and thoracolumbar/lumbar curves, with a curve magnitude difference less than 10 degrees°. The concave and convex sides of the paraspinal muscle region were identified based on ultrasound images obtained from the Scolioscan. The demographic data of the subjects are shown in Curve type in this study is defined on the basis of the guidelines from the Peking Union Medical College (PUMC) classification system . The subThe parameters of the sEMG assessment were formulated based on the Surface EMG for Noninvasive Assessment of Muscles (SENIAM) standards . The sEMDuring the collection of the sEMG data for habitual standing, the study participants were barefoot with arms relaxed and lightly clasped in front of their body and feet positioned 20 cm apart. They were instructed to focus straight ahead and look at a designated point . The habSuggested standing position: the head and ankles should be straight, shoulders and hip are level, kneecaps face the front, the head and knees are straight, and the chin should be parallel to the floor and aligned with the ears. The lower back should be slightly bent forward with the aid of the chest, stomach, and buttock muscles.Suggested sitting position: the head and ankles should be straight, shoulders and hips are level, kneecaps face the front, and the chin should be parallel to the floor and aligned with the ears. The lower back should be slightly bent forward to support the body with no extra weight distributed onto the spine.The requirements of the standing and sitting positions were used per recommendations in McKenzie  and CheuThe measurements of the EMG activity of the paraspinal muscles of the subjects were taken during the habitual postures and suggested positions of standing and sitting for a duration of 1 minute and repeated twice. A band pass filter that ranged from 10 to 500 Hz was applied to eliminate undesired artifacts, such as sudden movement, and a 60 Hz notch filter was used to eliminate noise. The sEMG signals were sampled at a rate of 2048 Hz. The EMG raw data were averaged by using root mean square (RMS) to obtain the average amplitude of the EMG signal. The RMS sEMG ratio of the subjects was calculated based on the following equation : (1)RMS The ratio is an index of the symmetric sEMG activity of the tested muscles, in which when the ratio is 1, the tested pairs of muscles have identical sEMG activity from the concave and convex sides of the tested muscle. If the ratio is less than 1, the concave side of the muscle has stronger sEMG activity than the convex side. If the ratio is larger than 1, the concave side of the muscle has weaker sEMG activity than the convex side. The equation was applied to assess the effectiveness of the suggested positions for the scoliosis subjects. The suggested positions are effective if the ratio is closer to 1 compared to the ratio recorded for the habitual postures.t-testing with the significance level at p < 0.05. Besides, the difference between habitual and the suggested standing and sitting sEMG ratios was compared through the significance of a one-sample t-test with a test value of 1. The level of significance is p < 0.05.Statistical analyses were conducted by using the SPSS 19 program for Windows. The difference between the convex and concave sides during habitual standing and sitting was compared by s) of the paraspinal muscles of the subjects during habitual standing and sitting and the suggested standing and sitting positions. The highlighted parts indicate posture improvement under the guidance of the physiotherapist, in which the RMS sEMG ratio of the suggested positions is closer to 1 as opposed to that obtained by the same habitual posture.p = 0.043). This is consistent with the findings of Mannion et al. where the concave side of the paraspinal muscles has lower bioelectric activity which caused muscle impairment at the convex side [Based on the acquired data in vex side .However, a contradictory situation is found in the PUMC type Ib subjects for most of the tested regions; the RMS EMG values show a different result than that for the PUMC type Ia and IIc subjects. The RMS EMG values at the muscle region on the concave side are larger than those on the convex side. This may be due to the comparatively smaller degree of spinal curvature of the PUMC type Ib subjects (mean = 13.6); therefore, the reduction of bioelectric activity on the concave side was not reflected from the sEMG data. It is important to note that the convex side where the spinal deformity has taken place does not necessarily incur stronger sEMG values as opposed to the concave side. An influencing factor to take into consideration could be due to the degree of the spinal curvature.The treatment for adolescents with mild idiopathic scoliosis is often passive, with only periodical observation by orthopaedists. It is possible that scoliotic adolescents suffer from progressive spinal curvature in a short period of time during puberty . AdolescDespite the suggested therapy in which patients with mild idiopathic scoliosis should be periodically observed rather than prescribed with any type of treatment or exercise during the early stages, unless a rapid change takes place in the curvature angle or there is spine rotation, adolescents with mild scoliosis could be treated with the use of exercise, as indicated by work in the literature; see –35. In 1In this study, the subjects are asked to perform habitual standing and sitting and given suggested standing and sitting positions, which are recorded with the EMG method. The suggested positions as guided by a physiotherapist with the aim of retaining postural balance of the subjects during standing and sitting are considered to be common daily positions. By restricting the scoliotic adolescents to a balanced posture, the paraspinal muscles between the two sides of the spine were able to achieve a more balanced state.The results in p = 0.023) while none of the tested muscle regions have a ratio with a significant difference with 1 during the suggested standing posture. During habitual sitting, the ratio for both the trapezius area and erector spinae at the thoracic region tends to have a significant difference of 1  while during the suggested sitting posture, only the ratio of the trapezius has a significant difference with 1 (p = 0.025). The results show that the subjects can achieve a more balanced sEMG signal during the suggested posture while standing and sitting.Overall, during habitual standing, the RMS sEMG ratio for the trapezius region has a significant difference with 1  and therefore reduce the possible need to prescribe brace wear treatment due to its associated psychological stress and negative impacts on adolescents. It is considered that, through motor learning of the suggested positions as a new type of motor task, the concave and convex sides of the paraspinal muscles will be more balanced.


A 21-year-old man with a history of left uncorrected cryptorchidism presented to the Emergency Department with sudden onset of left groin pain. He did not have any nausea, vomiting, or fevers. Additionally, there was no history of trauma, change in bowel habits or genitourinary symptoms prior to the onset of the pain. Physical examination revealed a tender, swollen and painful undescended testis located in the inguinal area with an empty ipsilateral hemiscrotum. Men presenting with pain and undescended testis are more likely to have testicular torsion, malignancy, associated inguinal hernia or trauma. The extra-scrotal position of the testis expands the differential diagnosis to also include suppurative inguinal lymphadenitis, fat necrosis, impacted ureteral stone or epididymitis. Doppler examination; however, was not performed in our patient so as to avoid unnecessary delay in the surgical exploration of the groin to potentially avert missed testicular torsion or incarcerated hernia. Intra-operative findings revealed torsion of the appendix testis in an inguinal cryptorchid testicle. The spermatic cord was untwisted and no associated hernia sac was identified. The necrotic appendix was excised and cryptorchidism corrected. Torsion of the appendix testis in an undescended testis is an uncommon condition. Although it can be managed conservatively, the diagnosis is significantly more challenging than in normally descended testes. Hence, it should reduce the threshold for surgical exploration to clear up any doubts. Excision of the twisted appendix can rapidly relieve refractory pain, avert reactive epididymitis as well as hasten recovery, which seems to have happened in our patient.


Solid organ transplantation has emerged as a life-saving treatment for many patients suffering from end-stage organ failure. Organs have been successfully recovered after a variety of aggressive interventions. We propose that decompressive laparotomy, when clinically indicated, should be considered in the aggressive resuscitation of potential organ donors. A thorough literature review examining aggressive interventions on potential organ donors was conducted after experience with a unique case at this institution. Articles were reviewed for the types of interventions performed as well as the time frame in relation to organ donation. In our case, several ethical issues were raised when considering decompressive laparotomy in a patient pronounced dead by neurologic criteria. We propose that having a surgical intensivist involved in the management of potential donors will further increase the salvage rate, as more invasive resuscitation options are possible. Solid organ transplantation has emerged as a life-saving treatment for many patients suffering from end-stage organ failure. While the volume of patients on the transplant waiting list continues to rise each year, the number of organs donated falls far short of the number needed. Currently, there are over 131,000 patients waiting for a solid organ transplant, and only 11,800 donors were recovered in 2013 . Here, wA 52-year-old male was admitted with a severe intracranial abscess and progressed to a diagnosis of death by neurological criteria. The local organ procurement organization (OPO) evaluated the patient for potential organ donation and obtained informed consent from the next of kin for organ donation and potential resuscitative interventions. Aggressive resuscitation was continued. Our center uses an organ donor intensivist (ODI) consult service model with medical, anesthesia, and surgical intensivists sharing ODI call coverage. The patient required vasopressors and intravenous fluid resuscitation. Several hours into his resuscitation, his abdomen became increasingly distended and firm. Peak airway pressures were elevated to 34 mmHg, and a bladder pressure obtained after administration of paralytic medication was elevated at 31 mmHg. Concerned for abdominal compartment syndrome, the ODI contacted the acute care surgical service for discussion of decompressive laparotomy. The attending surgeon on the acute care surgery service was a fellow ODI. Approximately 90 minutes after this diagnosis, the patient was taken to operating room where decompressive laparotomy was performed and a temporary abdominal closure device was placed. Peak airway pressures improved after operative intervention, and the patient returned to the intensive care unit for continued resuscitation. Later the next day, liver and kidneys were successfully procured for transplant and the liver was successfully transplanted. The recipient is reportedly doing well.Aggressive resuscitation of potential donors has increased the number of organs available for transplantation. Use of intensivists and structured resuscitation protocols has contributed to the improved recovery efforts . We propBecause the next of kin had already consented to organ donation, the OPO bears the costs of the procedure. Neither the donor's insurance company nor the donor's estate bears any of the associated costs.The informed consent process with the next of kin included an explanation of the possible procedures used to optimize organ donation potential, including invasive surgical procedures if necessary. This encompasses the decompressive laparotomy, but this procedure was not anticipated and therefore not specifically discussed with the donor's next of kin. The next of kin was not available prior to the procedure for additional discussions. When possible, informing the next of kin of this additional operative intervention outside of procurement is best in caring for the donor and respecting the family.In this case, the decompressive laparotomy can be justified because the potential benefit of organ preservation for donation outweighs the risks associated with the laparotomy. Since the potential donor is already dead, he could not be harmed in the sense of pain, morbidity, or mortality. Concerns about causing distress to the family and respecting the body's dignity are addressed by seeking the consent of the family and treating the body with respect, which includes only performing the procedures necessary and performing them with the same attention and consideration that we would do with a living patient. But the fact that the potential donor cannot suffer pain, morbidity, or mortality does not mean that any and all efforts would be justified. Exactly where the line should be drawn would depend on the details of a particular case: how strongly the potential donor and his next of kin wished for donation and what they are willing to consent to, how likely it is that the organs will be useable after the procedure, how aggressive and invasive the procedure is, whether the use of the operating room or other resources would make those resources unavailable to a living patient who needs them, and other such considerations.Medical and anesthesia intensivists are extremely beneficial in optimizing most resuscitation parameters but are limited in the ability to perform certain invasive procedures like laparotomy. This highlights the benefit of having surgical involvement in the organ donor management program. Acute care surgeons, specializing in trauma, emergency surgery, and critical care, are ideally suited to this supportive role for the organ donor management program in donor centers. Many potential donors present as a result of devastating traumatic injuries and represent a patient population extremely familiar to trauma and acute care surgeons. Isolated case reports detail success in using damage control laparotomy and decompressive laparotomy specifically in attempts to salvage organs; however, in our literature search, only one report of each was noted [With all procedures performed in the management of potential organ donors, ethical considerations are particularly important and sensitive. The key ethical issues are related to financial responsibility for the procedures, proper informed consent, and the ethical appropriateness of aggressive, invasive procedures in an already-dead donor .Because as noted , 4.In summary, this case represents an important example of aggressive treatment and resuscitation resulting in successful procurement of multiple organs for transplant. With the advent of expanded-donor criteria and the increasing urgency in need for organs, surgical salvage procedures should be considered in all potential organ donors. Decisions in these cases should be guided by a multidisciplinary team including organ procurement organizations and surgical intensivists.


Treatment with ritonavir-boosted protease inhibitors and nucleoside analogues frequently leads to rises in lipids, which might increase the cardiovascular risk. The aim of this study was to describe changes in lipid levels among HIV positive patients participating in the GARDEL study.The GARDEL study compared the efficacy and safety of a dual therapy (DT) combination of LPV/r 400/100 mg BID+3TC 150 mg BID to a triple therapy (TT) with LPV/r 400/100 mg BID+3TC or FTC and a third investigator-selected NRTI in fixed-dose combination among HIV+ treatment naïve patients. We compared changes in lipid levels from baseline to week 48 in both arms.Patient's characteristics were well balanced regarding mean baseline total cholesterol , triglycerides , LDL-C  and HDL-C . Changes in total cholesterol, LDL-C and HDL-C were higher in DT arm, compared to TT . Increase in triglycerides was higher in TT compared to DT (55% DT vs 92% TT) . In TT aChanges in lipid parameters were observed in both arms. Albeit the increase was numerically higher for cholesterol  in DT arm while TT arm had higher increases in TG; no difference was observed when week 48 values were compared with the NCEP ATP III goals for cardiovascular risk reduction . So, the


Lung immaturity due to preterm birth is a significant complication affecting neonatal health. Despite the detrimental effects of supplemental oxygen on alveolar formation, it remains an important treatment for infants with respiratory distress. Macrophages are traditionally associated with the propagation of inflammatory insults, however increased appreciation of their diversity has revealed essential functions in development and regeneration.Macrophage regulatory cytokine Colony-Stimulating Factor-1 (CSF-1) was investigated in a model of neonatal hyperoxia exposure, with the aim of promoting macrophages associated with alveologenesis to protect/rescue lung development and function. Neonatal mice were exposed to normoxia (21% oxygen) or hyperoxia ; and administered CSF-1  or vehicle (PBS) in two treatment regimes; 1) after hyperoxia from postnatal day (P)7-11, or 2) concurrently with five days of hyperoxia from P1-5. Lung structure, function and macrophages were assessed using alveolar morphometry, barometric whole-body plethysmography and flow cytometry.Seven days of hyperoxia resulted in an 18% decrease in body weight and perturbation of lung structure and function. In regime 1, growth restriction persisted in the Hyp + PBS and Hyp + CSF-1 groups, although perturbations in respiratory function were resolved by P35. CSF-1 increased CSF-1R+/F4/80+ macrophage number by 34% at P11 compared to Hyp + PBS, but was not associated with growth or lung structural rescue. In regime 2, five days of hyperoxia did not cause initial growth restriction in the Hyp + PBS and Hyp + CSF-1 groups, although body weight was decreased at P35 with CSF-1. CSF-1 was not associated with increased macrophages, or with functional perturbation in the adult. Overall, CSF-1 did not rescue the growth and lung defects associated with hyperoxia in this model; however, an increase in CSF-1R+ macrophages was not associated with an exacerbation of lung injury. The trophic functions of macrophages in lung development requires further elucidation in order to explore macrophage modulation as a strategy for promoting lung maturation. Immaturity of the lungs due to preterm birth is one of the most significant complications affecting neonatal mortality. As preterm infants with respiratory distress are born with lungs at an anatomical stage not yet conducive to gas exchange, routine care practices include mechanical ventilation and oxygen supplementation. However, these lifesaving interventions can also cause permanent damage to the developing lung, resulting in a chronic lung disease termed bronchopulmonary dysplasia (BPD) ,2. Moderin utero at around 36 weeks gestation, with 85% of alveoli formed after birth [Delivery of high concentrations of oxygen to neonatal animals is widely used to investigate the basis of arrested alveolar development associated with BPD in preterm infants ,6. In huer birth . In the er birth –11. UndeMacrophages are key immune cells commonly associated with inflammation and the propagation of tissue injury associated with oxygen toxicity in hyperoxia exposure. However, a greater appreciation of macrophage diversity is revealing that this heterogenous cell type is also essential in regulating organ development and regeneration –15. In pColony-Stimulating Factor-1 (CSF-1) is a key regulator of macrophage differentiation, proliferation, survival and activation, and acts as a pleiotropic growth factor essential in reproduction and organogenesis . During CSF-1 administration has been shown to enhance organogenesis. Increased branching morphogenesis was observed in embryonic kidney explants following the addition of CSF-1 , and in CSF-1-responsive macrophages in developing embryonic kidneys and lungs exhibit a gene expression profile characteristic of an M2 or tissue remodelling-type macrophage . SimilarThe aim of this study was to determine the impact of CSF-1 administration on lung maturation in a model of hyperoxia-associated lung developmental perturbation, when delivered either as a treatment after hyperoxia or when administered prophylactically with concurrent hyperoxia exposure. In particular, we assessed lung functional maturation and the impact of a CSF-1-mediated increase in macrophages. We found that CSF-1 did not rescue the growth and pulmonary defects associated with hyperoxia in this model; however the concomitant increased infiltration of CSF-1-responsive macrophages was not associated with exacerbation of lung injury.csf1r-EGFP mice, which express enhanced green fluorescent protein (EGFP) in cells of the myeloid lineage via the insertion of an EGFP transgene under the control of the Csf1r promoter [csf1r-EGFP mice were allowed to litter-down naturally in individually ventilated cages . Litters were randomly assigned to receive either normoxia  or hyperoxia (65% oxygen). Hyperoxia was achieved by mixing 100% medical grade oxygen and medical grade dry air, delivered to cages via Perspex(R) tubing into ventilation ports. Oxygen and carbon dioxide concentrations were continuously monitored by a gas analyser . Dams remained with their own pups for the entire experiment. Boxes were briefly disconnected from hyperoxia gases for a maximum of 15 minutes when injections were performed. Following hyperoxia exposure , mice were maintained in room air until experimental endpoints at P11 and P35.All animal experiments were approved by the Monash University Animal Ethics Committee and conducted in accordance with the “Australian Code of Practice for the Care and Use of Animals for Scientific Purposes”. This study utilised promoter . Pregnanin vitro culture assays demonstrating CSF-1-mediated proliferation of bone-marrow derived macrophages (data not shown). In treatment regime 1, neonatal pups were continuously exposed to hyperoxia from birth for 7 days with subsequent injections performed once daily for 5 days from P7-11. In treatment regime 2, newborn pups were exposed to hyperoxia and were administered CSF-1 concurrently, and therefore a shorter exposure of 5 days was used to facilitate 5 concurrent daily injections. At experimental endpoints  mice were humanely euthanised via cervical dislocation for morphometric or flow cytometric analysis.Hyperoxia-exposed littermate pups received either mouse recombinant CSF-1  or vehicle (PBS), administered via intraperitoneal (i.p.) injection at a final volume of 25 μl. Biological activity was confirmed with Respiratory function was assessed using unrestrained barometric whole-body plethysmography, as described previously ,32,33. Iin situ via intratracheal instillation of 10% buffered formalin at a pressure of 20 cmH20. The entire thorax was subsequently immersion fixed for 24 hr before lungs were dissected. The left lobe was processed, embedded in paraffin wax, sectioned at 5 μm, mounted on Polysine™ slides  and stained with haematoxylin and eosin. Images from 15 non-overlapping fields of view were captured at × 400 magnification and ImagePro Plus software  was used to assess mean linear intercept (MLI), as an estimate of alveolar diameter, and tissue/airspace percentage, as described previously [For morphometric assessments, the trachea was cannulated immediately after euthanasia and the lungs inflation-fixed eviously . In brie6 cells were immunolabelled with anti-CD45 APC Cy7-conjugated  and anti-F4/80 APC-conjugated  antibodies. Samples were run on a BD FACSCanto II cytometer (BD Biosciences) and data analysis was performed using FlowJo FCS analysis software .To assess macrophage populations in hyperoxic lungs and their response to CSF-1 treatment, whole lungs of littermate mice underwent flow cytometric analysis at the end of treatment at P11 or P5, with the final injection being administered 3 hours before lungs were collected. Whole lungs underwent enzymatic and mechanical digestion to yield a single cell suspension as described previously . Red bloThis study included 17 litters of mice, ranging between 4 and 8 pups per litter. The N numbers for bodyweight and lung function analysis outlined in appropriate figure legends are a cumulative of a minimum two repeated experiments. Flow cytometric analysis was performed on one representative litter for inter-litter comparison. Data are presented as mean ± standard error of the mean (SEM). Graphing and statistical analyses were performed using GraphPad Prism™ . Specific statistical tests used are outlined in figure legends. A p value of less than 0.05 was considered statistically significant.Neonatal murine hyperoxia is a model of impaired alveologenesis and a clinically relevant model of preterm lung damage. In our hyperoxia model, neonatal mice were exposed to 65% oxygen from birth for 7 days. Hyperoxia significantly decreased growth of neonatal mice. At birth there was no significant difference in body weight between groups (p = 0.09); however after 7 days of hyperoxia exposure body weight was decreased by 18% compared to normoxia-exposed offspring  to adulthood at P35 (21.17 ± 0.47 g vs. 17.70 ± 0.76 g, p < 0.001; Figure Removal of mice from hyperoxia resulted in a marked improvement in lung structure. At the end of treatment at P11, there was no significant difference in MLI, percentage tissue or percentage airspace amongst any of the three groups Table . HoweverA time course analysis of lung function was performed from the conclusion of hyperoxia exposure at P7, at the end of treatment at P11, and weekly throughout development from P14 to adulthood at P35 (Figure 5 vs. 144 ± 3.1 × 105, p < 0.05; Figure csf1r-EGFP mice, which express EGFP under control of the csf1r promoter, identified CSF-1-responsive cells. The number of CSF-1R+ cells significantly increased by 34% with CSF-1 treatment compared to PBS (16.7 ± 0.2 × 104 vs. 22.2 ± 2.2 × 104, p < 0.05; Figure 3 vs. 118 ± 9.0 × 103, p < 0.05; Figure To investigate the impact of CSF-1 treatment on lung macrophages, flow cytometry was performed on littermate hyperoxic lungs after treatment with either PBS or CSF-1 at P11 Figure . The totIn the second treatment regime, CSF-1 was administered in conjunction with high oxygen exposure to assess the potential of CSF-1 to mitigate lung damage and promote alveolar formation. In this more clinically relevant regime, neonatal mice were exposed to hyperoxia for 5 days to allow for 5 concurrent daily injections of CSF-1 or PBS. Body weight was measured to assess growth over the time course of development, from the end of treatment at P5, and weekly throughout postnatal development from P14 to adulthood at P35 (Figure The effect of hyperoxia and concurrent CSF-1 treatment on structural development of the lung was assessed at P5 Table . No signPlethysmographic analysis of breath parameters Figure  indicateCsf1r-EGFP transgene expression on CD45+ leukocytes revealed that the number and percentage of CSF-1R+ cells within the lung was comparable amongst both PBS and CSF-1-treated mice (Figure To investigate the impact of CSF-1 treatment on lung macrophages, flow cytometry was performed on littermate lungs at P5, after either PBS or CSF-1 treatment was delivered concurrently with hyperoxia. Total lung cell number was not altered with CSF-1 treatment, with no significant change compared to PBS was observed Figure B. Csf1r-Oxygen supplementation remains a critical and lifesaving intervention for infants with respiratory distress; however, it can result in detrimental alterations to the developing lung. Exposing neonatal mice to high concentrations of oxygen mimics the damage to the lung associated with BPD, providing a relevant model to assess the function of CSF-1 in rescuing and enhancing alveolar formation. This study employed a hyperoxia regime of 65% oxygen for 1 week after birth, providing a milder, more clinically-relevant exposure that more closely resembles the oxygen therapy regime of ventilated premature infants than other previous studies . It was Functional perturbations were also revealed following hyperoxia. Unrestrained barometric whole-body plethysmography proved a beneficial analytical technique sensitive enough to reveal measurable differences in neonates at 7 days of age. While the accuracy of absolute quantitation in small animals is debatable , this teHarnessing the organogenic importance of growth factors involved in alveolar formation has been a fundamental strategy for promoting lung maturation. Factors such as keratinocyte growth factor , hepatocCSF-1 provides a particularly attractive candidate for use in this setting because it is a growth factor with proven organogenic ,28, antiThe role of CSF-1 in polarising macrophages towards an M2 phenotype may also provide beneficial effects in hyperoxia through its involvement in immune dampening . InflammThe infiltration of inflammatory macrophages is associated with the progression of lung injury and pathology ,61. HoweIn the present study, CSF-1 was administered to a neonatal murine hyperoxia model and growth, lung structure and respiratory function were assessed. In the first treatment regime, CSF-1 was administered post-hyperoxic injury to investigate whether CSF-1 could rescue developmental perturbation and promote alveolar formation. Results demonstrated limited success of CSF-1 in the mitigation of hyperoxia-induced injury. Deficits in growth were not improved, with the body weights of CSF-1-treated mice comparable to the Hyp + PBS group. With regard to lung function, a negative impact was revealed at P11 at the end of treatment. In parameters such as total cycle time, expiration time and minute volume/body weight ratio the detrimental effect of hyperoxia was exacerbated with CSF-1. Interestingly, these initial defects did normalise over time and by P35 no differences between treatment groups were observed. This normalisation was also seen in the Hyp + PBS group, and therefore CSF-1 treatment did not accelerate or enhance functional recovery, although increased numbers of CSF-1R+ macrophages were evident. Nevertheless, it is noteworthy that CSF-1 did not have a negative impact on the parameters measured in the adult. It is interesting that earlier perturbations observed with hyperoxia resolved by adulthood. Whether this impacts on the lungs capacity to cope with challenges and aging in later life is unknown. Furthermore questions remain about potential negative effects associated with catch up growth. Indeed CSF-1R+ macrophages have been shown to be increased in the alveolarisation stage of lung development , howeverOxygen supplementation is a critical life saving intervention for babies with respiratory distress, and clinically any maturation-based therapies would need to be delivered concurrently. Therefore, in the second part of this study, CSF-1 was administered from birth in conjunction with hyperoxic exposure, in a setting where a prophylactic effect of CSF-1 treatment was aimed at preventing damage and alveolar loss and enhancing alveolarisation. A shortened exposure time was utilised to enable the 5 daily doses to be administered in conjunction with hyperoxia. The lack of growth restriction is likely due to the decreased oxygen exposure, as pup litters size was consistent amongst regimes. Structurally and functionally, the lungs of both hyperoxia groups were impacted negatively. Morphometric analysis in the adult revealed significant increases in MLI, with fewer, larger alveoli in both the Hyp + PBS and Hyp + CSF-1 groups compared to Norm. Although CSF-1 was not found to improve the structural alterations caused by hyperoxia, this treatment did not exacerbate the negative impact of hyperoxia and no significant difference between PBS and CSF-1-treated mice was observed. There were no differences between any groups at P5, although this time coincides with the beginning of the alveolarisation phase. Of note is that Hyp + CSF-1 values began to approach Norm values in all parameters. Functionally in the adult, CSF-1 did not exacerbate the adverse effects on lung function that were characteristic of hyperoxia exposure, as represented by the Hyp + PBS group. Although no negative effect of CSF-1 was identified, it must be noted that no functionally beneficial effects with regard to preventing hyperoxic damage or further enhancing lung development were observed. The only changes with CSF-1 at P35 were in tidal volume and minute volume. However, these were normalised when body weight was accounted for and, relative to their size, there were no changes in these parameters as a result of CSF-1 treatment.Overall, CSF-1 treatment showed little effect in promoting alveolar formation in the hyperoxic settings employed in this study. Ongoing studies examining the role of M2 polarisation in the protection and rescue of hyperoxic injury with be important before a definitive statement that CSF-1-responsive macrophages are not effective can be made. Also further experiments utilizing a premature birth model – where animals are prematurely removed from their high uterine CSF-1 environment – will be important to better model this aspect of the clinical situation. However, in this study the finding that CSF-1 treatment did not exacerbate the pathological response to high oxygen levels was encouraging in that macrophage-mediation may provide a novel strategy for the immunomodulation and trophic promotion of alveolar development. In particular in the second regime where potential negative outcomes were a legitimate concern, CSF-1 did not exacerbate damage with concurrent oxygen supplementation. This is a significant finding that is important for any potential clinical use of CSF-1, and supports ongoing studies to improve understanding of the role of CSF-1 in alveolar formation.


Different methods for dressing of donor site of skin graft in burn patients have similarly pain, limitation of mobility of donor site and local complications such as infection and scar. Amniotic membrane has used for improvement of healing in some wounds. Accordingly in this study amnion was used as biologic dressing for donor site of skin graft to evaluate it’s efficacy in improvement of pain, move score and the risk of local infection. Study was done as clinical trial over 32 admitted patients in burn department of Beasat hospital. Amnion was prepared in elective caesarean section after rule out any placental site for risk of torch and viral infection. Skin graft was taken from two sites in every patient. One site dressed with amnion and another with routine dressing. Then two sites were compared about severity of pain, move score, infection and time of dressing sloughing.Fourteen patients were women and 18 men. Mean score of pain and movement up to fourth and fifth post operative day respectively was less than control site. No difference is seen about infection and dressing slough in two sites. It seems use of amnion for dressing of donor site probably cause rapid epithelialisation and wound healing and can improve pain and move score in early post operative days. Accordingly it is expected to need less analgesia and low rate of immobilization and following complications and earlier discharge of patients. Therefore the present study was carried out with an aim to reveal the efficiency of the biological dressing with amniotic membrane for the reduction of pain in the donor site, improvement of the body movement in the donor site limb and evaluation of duration of dressing sloughing from the wound and risk of infection. Annually many individuals in low and middle income countries suffer from burn injuries.This study was a clinical trial performed on 32 patients. These patients were hospitalized in the burn ward of the Besat Hospital due to burns and among the patients, the individuals aged over 18 years were enrolled. Each patient was considered as an intervention group and also as a control group. The one side of an organ was considered as an intervention area and the other side as a control region. The amniotic membrane in a fresh form was provided from the Department of Obstetrics and Gynaecology of Fatimane Hospital in an elective caesarean section. Infections to TORCH, hepatitis B, hepatitis C, AIDS, and syphilis before childbirth was not included. For hepatitis B and C, AIDS and syphilis infections, serological tests of HIVAb, HBS Ag, HCVAb and VDRL were carried out. After the cesarean section and under sterile conditions, the amniotic membrane was separated and placed in the normal saline solution and was immediately transferred to the burn department of the Apostolate Hospital in less than 6 hours from its isolation to be used for the study. The donor from the two limbs and or two harmonious body sections was selected using an electric dermatome with the thickness of 0.014 inch from each side. The dressing of the donor site in one side was done with vaseline gauze and was bandaged. For the dressing of the donor site on the opposite, the amniotic membrane was placed on the donor site and then the vaseline gauze on the membrane and then the dry gas was introduced and bandaged. The limbs that were subjected to study (i.e. the amniotic membrane placed on the donor site) was coded A, and the limb which was regarded as the control site  was coded B. After the 24 hours of surgery, the bandage and the dry gases of both sites (case study and control) were removed till the level of vaseline gauze and the two sites were exposed to open air  and later, the pain score, easiness in the move of organs, the spontaneous separation time of the last layer of dressing on the donor site and the condition of sepsis according to the protocol was provided. The pain score and easiness in the movement of the limbs that considered as the donor sites were determined by patient with the 1-5 and 1-10 score respectively in a daily examination from the patients and recorded in a questionnaire. Score 1 represented the lowest pain level and organ movement and 5 showed the highest organ movement level and 10 demonstrated the highest pain level and the validity of the infections in the two sites of case and control based on the daily clinical examination in the case of clinical symptoms based on the probability of sepsis , the cultivation was procured and in the case of bacterial growth over 100,000 colonies, it was considered positive. The patients who were discharged early post operatively, they were followed in the Sheikholraeis clinic and the required information about pain, organ movement, separation of dressings and sepsis were recorded in a questionnaire. The collected information from patients was eventually entered in the SPSS software  and was analyzed using Mann Whitney test. In the context of investigating the presence of pain difference in various days in the donor sites, t-test was used till the fifth day and Mann Whitney test from the sixth day for the comparison of pain and easiness of the limbs movement scores.th to 14th day, there was no significant difference in the pain score between two sides, although in all these cases, the pain score in the intervention area was less than the control area. The status of movement range in the intervention and control areas were demonstrated in Thirty two patients with range of 18-88 years were enrolled while 18 patients (56%) were male. The pain status in the intervention and control areas were shown in p=0.01) showing that the intervention cases which were dressed by the amniotic membrane had a better movement status in comparison to the routine dressing. From the 6th day to the 14th day, there was no significant difference between the movement of limbs in both sides, although in most of the cases, the intervention patients had a better movement status in relation to the control organ. The movement condition in the intervention limbs till the fifth day of hospitalization in comparison to the control limb exhibited a significant difference (p=0.29). Regarding the infection of intervention and control group, it was clear that two infection cases had appeared on the site dressed with amniotic membrane where in the general dressing site (control area), no infection was observed. The difference for infection in the dressings of intervention and control organs was not significant (p=0.49).The dressing separation time in the intervention cases from the days 3 to 18 after the surgery was about 8.15±3.4 days and in the control cases, the dressing separation in the days 5 to 21 was about 9.2±3.7 days while the difference of the dressing separation in intervention and control organs was not significant , and respiratory, gastrointestinal, endocrine and electrolyte disorders.20 Based on these complications after surgery especially skin transplantation, it is of great importance to prevent and reduce the complications few days after surgery. The coverage of donor site requires materials that preserve the epidermal function and integrate itself into the process of healing.16After skin grafts, the donor site was painful and caused immobility of patients and also prone them to infection, hypertrophic scar formation and changes in color21 All biologic dressings are susceptible to early reaction and the only exception is the amniotic membrane.21 The use of amniotic membrane as biological dressing in the treatment of extensive burn wounds has been described in order to early recovery of patient, improvement of wound healing and its quality.15 Studies demonstrated that use of amnion in burn wounds can lead to reduction of pain intensity and prevent water and electrolyte disturbances and also can help for early preparation of the wound bed for grafting.20,21Biological dressing was introduced as a gold standard for temporary covering of wounds.16The preparation and application of amniotic membrane according to our proposed method performed with simplicity and efficiency, lead to a significant difference for 3 variables between donor site with routine dressing and dressing with amniotic membrane. Biologic dressing lead to significant reduction of pain score at the site of skin graft in the first few days after surgery. In fact with covering of donor site with amnion, we introduced an occlusive dressing. Growth factors extravasated and accumulated in the interface of biologic dressing and abraded epidermis. This moisture and suitable interface could hasten the wound healing process and epithelialization.4Amnion prevents the wound from irritation, evaporation, dryness and also nerve ending stimuli on the wound surface. These characters could not be seen in routine dressings. In the first 24-48 hours after any skin abrasion, skin harvesting or even primary repair in surgical wounds, basal epithelial cells can cause a water tight closure in wound layer together with migration and mitosis processes.16This layer does not have enough strength against external mechanical stresses and is fragile. With disruption of this fine neo-epithelium, underlying nerve endings are exposed and suffered with contact to non-biological dressings. But after fifth day of graft taking, epithelialization reaches to optimal thickness and resistance in order to preserve its integrity against external stress. Therefore, pain score of both studied and control site did not show any difference after this time. Less pain at the first few days of skin harvesting is important. Patients could be mobilized soon and the need for analgesia during hospitalization is decreased and subsequent complications in respiratory and coagulation system are prevented.16Early and feasible mobility of patients with application of amnion over the donor site seems to be a secondary effect of its pain subsiding. Although in this study, there was not statistical difference about the time of dressing separation for two types of dressing, but clinically the mean time of dressing sloughing in studied limb was 8.5 and for routine dressing was 9.2 days. This clinical difference is due to improvement of epithelialisation with amnion dressing while amniotic membrane has low antigenicity.16 In this study, we did not have significant wound infection following to dressing by amnion. Amniotic membrane can prevent bacterial invasion to the donor site of wound by use of amniotic membrane. Similar conclusions could be seen in other studies.17,22Application of amnion in the first few days adheres tightly to wound surface and has positive modulation for wound healing process either in quality or in rapidity. Accordingly in this condition, possible morbidities of wound healing such as color change or hypertrophic scar would formation can be decreased. Dermal substitutes as biologic dressings can be used for wound care in plastic surgeries for this purpose but may not be cost-benefit.Reduction of pain severity and improvement of mobility with application of amniotic membrane over the donor site can cause earlier re-harvesting of graft from previous donor site specially in extensive burns and also decrease the duration of hospitalization, need for analgesia and complications of immobilization. Dressing of donor site of skin graft with amniotic membrane can reduce pain in the early post-operative days and facilitate the patient mobility and early sloughing of dressing from the wound bed. Due to ease of preparation and use of amniotic membrane as well as absence of adverse effects of its application, it is recommended to use amniotic membrane in the site of skin graft in order to be harvested in burn patients specially with high percentage of burn lesions for pain reduction and improvement of patient mobility and also acceleration of recovery of donor site for graft re-harvesting without any fear of infection.


In a cohort of 515 healthy stem cell donors we attempted to reproduce the correlation of the CXCL12 3’UTR SNP and mobilization responses and tested the role of miR941 in this context. The SNP was distributed with the expected frequency. Mobilization efficiency for CD34+ cells in WT, heterozygous and homozygous SNP individuals was indistinguishable, even after controlling for gender. miR941 expression in non-hematopoietic bone marrow cells was undetectable and miR941 did not interact with the 3’ UTR of CXCL12. Proposed effects of the SNP rs1801157 on G-CSF responsiveness cannot be confirmed in a larger cohort.The genetics responsible for the inter-individually variable G-CSF responsiveness remain elusive. A single nucleotide polymorphism (SNP) in the 3’UTR of CXCL12, rs1801157, was implicated in X4-tropic HiV susceptibility and later, in two small studies, in G-CSR responsiveness in patients and donors. The position of the SNP in the 3’UTR together with  Allogeneic G-CSF mobilized peripheral blood stem/progenitor cells from healthy volunteer donors have become the source of choice for allogeneic “stem cell” transplantation.–3 A hundhttp://www.targetscan.org/) as a putative binding site, although this interaction was not predicted by any other microRNA web-tools. The 4th base of this sequence being the polymorphic one, loss of the miRNA941 binding site of the SNP variant was predicted  stroma cells and differential interaction by miR941 with the WT- and SNP-variant of the CXCL12 3’UTR.th position of the seed by site directed mutagenesis , to match the sequence of the SNP) into plasmid pSUPER.retro.puro  and cloning of the whole 3’UTR of WT or SNP CXCL12ß (3285 bp) into the dual-luciferase plasmid pmirGLO . Plasmids were co-expressed in HEK 293 cells via lipofectamine . Transfection efficiency and expression of miR941 were tested by TaqMan qPCR (Applied Biosystems). Firefly and renilla luciferase activity were measured with the Dual-Glo Luciferase Assay System (Promega). In addition to miR941 expression by p.SUPER.retro.puro, synthetic miR941 and SNP-miR941 (Qiagen) were alternatively co-transfected with the pmirGLO 3’UTR variants. Cross-over analyses for WT- or SNP-3’UTR variant and WT- or mutated miR941were performed. Experiments were performed with commercial kits according to manufacturers’ instructions. Custom oligonucleotides are listed in Healthy volunteer stem cell donors were subjected to G-CSF induced stem cell mobilization as described (7.5–10 μg/kgBW*d in two divided doses q12h),–20 in prth day of G-CSF was 97.0±2.4 CD34+ cells/μl (mean±SEM), in agreement with published data for split-dose G-CSF.[These studies had been triggered by visibly published reports of SNP rs1801157 effects on CXCL12 protein levels and, likely in consequence thereof, on infectivity of X4-tropic HiV strains.–16 Thesese G-CSF. Distribuse G-CSF.–24 Chemose G-CSF. Moreoverse G-CSF. claimed se G-CSF. such as se G-CSF. but thishttp://www.targetscan.org/) in the region of SNP rs1801157 and loss thereof in the SNP variant was predicted. Even though none of several other commonly used miRNA search engines  predicted this interaction, to end the discussions about putative roles of SNP rs1801157 in mobilization responses, we explored whether the CXCL12 3’UTR polymorphism quantitatively affects mRNA expression and what is the role, if any, of miR941 in this context. For miR941 to differentially influence CXCL12 expression depending on the CXCL12 3’UTR genotype, the expectation would be that it must be expressed in BM stroma cells, the main source of CXCL12 in BM. miRNA expression arrays in non-hematopoietic BM cells, corroborated by qPCR, detected several highly expressed miRNAs, but no expression of miR941 (in silico analyses had suggested. Direct evidence was sought using dual luciferase assays. As we are showing, luciferase expression was not affected by co-expression of miR941, nor were differential effects on the different genotypes or differential effects of miR941 and mutant-miR941 observed (A putative binding site for hsa-miR-941 was predicted by Target Scan Human (f miR941 . We alsoobserved . LinkageIn summary, our studies definitively contradict previous reports about effects of the SNP rs1801157 on G-CSF responsiveness in donors, just as previous reports of an effect of this polymorphism on susceptibility towards X4-tropic HI virus were unhinged when sufficiently large cohorts of HIV patients were analyzed. We propo


The German transplantation system is in a crisis due to a lack of donor organs. Information campaigns are one of the main approaches to increase organ donation rates. Since 2012, German health insurance funds are obliged by law to inform their members about organ donation. We raised the hypothesis: The willingness to sign a donor card rises due to the subsequent increase of specific knowledge by receiving the information material of the health insurance funds.The objective of the study was to assess the influence of information campaigns on the specific knowledge and the willingness to donate organs.We conducted an online survey based on recruitment via Facebook groups, advertisements using the snowball effect, and on mailing lists of medical faculties in Germany. Besides the demographic data, the willingness to hold an organ donor card was investigated. Specific knowledge regarding transplantation was explored using five factual questions resulting in a specific knowledge score.P<.001), but not with the general education level (P=.155). Receiving information material was related to holding a donor card (P<.001), but not to a relevant increase in specific knowledge . The specific knowledge score and the percentage of organ donor card holders showed a linear association (P<.001).We recruited a total of 2484 participants, of which 32.7% (300/917) had received information material. Mean age was 29.9 . There were 65.81% (1594/2422) of the participants that were female. The mean knowledge score was 3.28 of a possible 5.00 . Holding a donor card was associated with specific knowledge (The information campaign was not associated with a relevant increase in specific knowledge, but with an increased rate in organ donor card holders. This effect is most likely related to the feeling of being informed, together with an easy access to the organ donor card. The German transplantation system is in a crisis due to a lack of donor organs. About 12,000 patients are waiting for an organ graft . Furthermore, information campaigns comparable to the campaigns of the health insurance funds should be repeated periodically. In addition, information about organ donation should be provided in more ways, as lessons in school, brochures in public buildings, or in television shows. Moreover, the access to organ donor card forms should be improved. These cards should be displayed at public buildings and additionally sent to every household every few years.