Computed microtomography, confocal microscopy, and histometry showed similarity between the PEO and SLA surfaces, with a trend toward the superiority of PEO in OVX animals. Thus, PEO surfaces were shown to be promising for enhancing peri-implant bone repair in ovariectomized rats.Plasma electrolytic oxidation (PEO) has been a promising surface coating with better mechanical and antimicrobial parameters comparing to conventional treatment surfaces. This study evaluated the peri-implant bone repair using (PEO) surface coatings compared with sandblasted acid (SLA) treatment. For this purpose, 44 Wistar rats were ovariectomized  or underwent simulated surgery  and received implants in the tibia with each of the surface coatings. The peri-implant bone subsequently underwent molecular, microstructural, bone turnover, and histometric analysis. Real-time PCR showed a higher expression of osteoprotegerin (OPG), receptor activator of nuclear kappa-B ligand (RANKL), and osteocalcin (OC) proteins in the SLA/OVX and PEO/SS groups ( Since the use of titanium emerged in the manufacture of medical, dental implants, and osseointegration principles, several modifications to this material have appeared ,2,3,4,5.Combining the favorable characteristics of microstructural and biological behavioral aspects is a concern related to this range of texturing options ,4,5, becThe techniques associated with biomimetic coatings, which incorporates calcium (Ca), phosphorus (P), and hydroxyapatite, ,13 have This electrochemical process gives titanium or its alloys mechanical and thermal superiority in addition to corrosive and tribocorrosive properties ,17,18,19Bone tissue quality is an essential factor in osseointegration, in which the implant and peri-implant bone complex undergo constant thermal, chemical, and mechanical oscillations throughout a coupled remodeling process . When a In addition, osteoporosis causes a significant decrease in bone density, and although it is not an absolute contraindication to implant therapy, it reduces implant survival rates ,24,25. TTherefore, this study aimed to evaluate a new dental implant coating method using PEO and calcium and phosphate ion aggregation in osteoporotic animals.Ti-6Al-4V implants  with machined surfaces and measuring 2 mm in diameter and 6 mm in length received two different implant surface topography treatments: PEO and sandblasted acid (SLA).in vitro studies ) was defined. This area was defined as the total area (0.25-mm margin around the implants: 2.38 mm × 2.38 mm). The CTAn software analyzes and measures images in grayscale (threshold). After adjusting and removing shades of gray from the corresponding area of the implant, the threshold used in the analysis was 45–255 shades of gray, which allowed us to obtain the volume of bone formed around the implants.Therefore, following the volumetric parameters suggested by the American Academy of Mineral Bone Research, the parameters related to the amount of bone tissue  and bone tissue quality (Tb.Th = bone trabecular thickness) were obtained. Tb.SP = separation of bone trabeculae, Tb.N = number of trabeculae, and Po. (Tot) = percentage of total porosity.® resin  until use. Resin was only used as a dipping medium. The samples were embedded in the resin, which was light-cured, and the Exakt processing protocol  was performed. Parts were sectioned and worn by using an automatic polishing cutting system  until a section approximately 100 μm thick was obtained.After the microtomography, the pieces were returned to the end of the laboratory that processed the calcified tissues. They were subjected to dehydration with a gradually increasing sequence of concentrations of alcohol of 70%, 80%, 90%, 95%, and 100%, and the solution was changed every 3 days. The dehydrated parts were placed in an orbital shaker  every day for 4 min. At the end of the dehydration step, the samples were immersed in a mixture of 100% alcohol and submitted to light-curing Techno Vit2 in size and corresponded to optical sections of 512 × 512 pixels. Thereafter, 2-μm sections were scanned for 2.5 min. Thus, 28 cuts were obtained for each 56-μm scan. The barrier filters used were BP 530/30 nm and 590 LP, combined with the activation of the double dichroic 488/568 nm. The photomultiplier was adjusted to 534 for calcein and 357 for alizarin. They were located under the microscope, and when light from the mercury lamp passed through these barriers and reached the specimen cuts, it was possible to visualize the fluorochrome colors. The blue filter facilitated the visualization of calcein, and the green filter facilitated the visualization of alizarin.The slides obtained were submitted to analysis using Leica CTR 4000 CS SPE laser confocal microscopy . These images were 1 × 1 mm2. The same procedure was performed for alizarin to obtain data on peri-implant bone dynamics. Using this methodological approach, bone turnover was represented by the difference between old bone (green) and new bone (red). In addition, the mineral apposition rate (MAR) was calculated by overlapping the images (red and green) and using the Image J® straight tool to measure the distance in μm from the beginning of the calcein precipitation to the other extreme of the alizarin precipitation.The tibial bone (peri-implant region) had two fluorochrome overlays ; each overlap represented a calcium precipitation according to each time interval (14 and 42 days), showing the conversion of old bone to new bone. These images were saved in TIFF format and analyzed using the color threshold tool in Image J . Each image was standardized according to hue, saturation, and brightness. First, calcein was highlighted, and the “measure” tool was used to provide the area in μm2. For the NBF calculation, the freehand selection tool was used to measure the area of newly formed bone in the region corresponding to the most central thread of the implant in pixels [After analysis by confocal microscopy, the pieces were washed in deionized water and stained with alizarin red and Stevenel blue. Histometrically, we calculated the linear extent of contact between the newly formed bone tissue and the implant surface (BIC) bilaterally and determined the newly formed bone area (NBF) in the most central thread on each side of the implant. Since each implant on the slide was a unit of analysis, readings were taken on both sides of the implant, so its mean would be representative of the unit. Thus, after photomicrography of the histological slides, which were saved as TIFF files, these were analyzed using Image J software . Using the straight tool, we calculated the bone perimeter formed at the BIC in pixelsn pixels .p < 0.05, using the statistical program SigmaPlot 12.0 . For the data obtained by microtomography, which included BV.TV, Tb.Th, Tb.N, Tb.Sp, and Po. (Tot), separate t-test statistical analyses were performed for the SS and OVX groups to enable the intragroup analysis to be performed. The analysis of peri-implant bone turnover via calcein and alizarin labeling was performed using a one-way ANOVA and Kruskal–Wallis one-factor tests, and the comparison of fluorochromes  was performed using a two-way ANOVA. The histometry results for Bone Implant Contact (BIC) and New Bone Formed (NBF) were analyzed with one-way ANOVA tests.The results were submitted to the normality test (Shapiro–Wilk), with a significance value of The sample size was determined according to the previously published manuscript , in whicp < 0.05; t-test).The results of ELISA test indicated a wide difference between the groups, with estrogen dosage less than 10 ng/dl in the OVX group and approximately 60 ng/dl in the SS group (The cellular responses of peri-implant bone tissue were compared by considering the SLA and PEO surface interactions (SS and OVX).p < 0.05). For the SS group, the highest values were for the PEO group (p < 0.05)  .The 3D representative images from the tomographic sections of each experimental group at 60 days postoperative may be seen in p = 0.726) and OVX (p = 0.428) groups, but PEO showed numerical superiority in low-density bone (OVX)  A.p = 0.350) and OVX/SLA groups (p = 0.428), there was no significant difference between groups. Complementary data showed that in the ovariectomized rats (OVX), the PEO surface tended to reduce bone porosity (Although Po. (Tot) values were numerically higher in the SS/PEO (porosity B.p > 0.05)  .Photomicrographs representative of fluorochromes calcein and alizarin red were showed in p = 0.299) and OVX (p = 0.614) groups. Alizarin labeling was also similar in the SS (p = 0.685) and OVX (p = 0.899) groups when compared with the SLA and PEO surface coatings  and OVX (p = 0.624).When comparing calcein and alizarin in the experimental groups, no statistical significance was found for the SLA and PEO surface coatings: SS (p = 0.795), and there was a tendency to higher MAR in the SS/PEO group when compared with the SS/SLA group (p = 0.211).Relative to the daily MAR, the SLA and PEO surface coatings were similar in the OVX group , with statistical results similar to those of the SLA surface in the SS groups . The analysis of the linear BIC was similar when the SLA and PEO surfaces were compared in the SS (p = 0.758) and OVX (p = 0.374) groups using the one-way ANOVA test  values showed no statistical differences in the coatings of experimental intragroup surfaces. However, in the OVX experimental model, the PEO surface presented a higher percentage of bone volume and less porosity than the SLA surface. Relative to bone quality, Tb.Th, Tb.Sp, and Tb.N were similar between groups.The PEO and SLA surface coatings tested in this study demonstrated great ability to promote bone formation irrespective of the experimental bone types in the BIC and in the NBF between the turns of the implant. He et al. demonstrated that the incorporation of Ca, P, or zinc into the surface coating, through PEO, can accelerate bone formation and remodeling and shorten the osseointegration period with better bone-bonding strength .Surface topography changes in osseointegrated implants aim to improve their mechanical properties and corrosion resistance in biological fluids, as well as the reparative biological responses of bone tissue. Similar to other materials used in physiological conditions, implant exposure to mechanical and biological factors can impair their survival and treatment success .Previous studies of PEO surface characterization have shown that on surfaces coated by this method, an oxide layer is produced, which provides increased wear and corrosion resistance, thermal protection, and the possibility of good adhesion by ions that are important to surface osseointegration, such as Ca and P. This incorporation of Ca and P showed a more homogeneous crystalline structure, as well as large, volcano-like pores, as proven by Scanning electron microscopy. It has also promoted antibacterial properties in microbiological tests ,2,7,21. In our study, the bone tissue dynamics represented by bone turnover and observed through fluorochromes were similar in the experimental coating surface groups. The PEO surface showed a better MAR trend in the SS group when compared with the SLA surface. For the OVX group, the PEO surface showed a tendency to higher MAR than SLA surface, and no statistically significant difference was found between the groups. However, the OVX animals in this study had slower bone turnover due to the non-systemic treatment of poor bone quality.Nevertheless, the values found in the PCR analysis of the present study indicated that PEO facilitated advanced bone repair in the OVX group, because at 42 days, PCR showed a reduction in OPG, RANKL, and OC proteins for this group. Thus, it is likely that osteocalcin expression had already occurred as part of the bone repair process during this period in the PEO/OVX group and that this was confirmed by microtomographic and histometric findings at 60 days, which showed parity with and slight superiority of the PEO surface over the SLA surface.The PEO surface method favors the addition of Mg, Al, Ti, Ca, and P molecules by providing improved protection against corrosion and wear, and the initial cellular response is higher with Ca and P, in relation to the others, due to their greater biocompatibility and bioactivity . AccordiThe study of implant bioactivity in low-quality bone tissue, as performed in this research, is not related to the treatment of osteoporosis through medications that reverse this condition, such as anti-resorptive and selective estrogen receptor modulators (SERMs). Previous studies have shown that systemic treatment assisted in the molecular responses of bone tissue biology ,44. ThusHowever, for medical and dental implant applications, characteristics other than bone formation should be observed to ensure optimal material performance and longevity. Thus, the PEO surface coating was shown to be extremely promising and offered excellent performance in terms of bone repair, as demonstrated in the present study. This surface coating also has superior mechanical ,17,18 anRegarding the limitations of the study, some analysis is very important to ensure the safety of the dental implants related to microgeometry, especially the first times of the assessment and the loading conditions. Future studies should investigate these coating surfaces in the first weeks of postoperative and also develop an experimental model to simulate the loading since the early stages of the osseointegration.Therefore, we concluded that both surface coatings demonstrated good performance in peri-implant repair in rats with low-density bones by improving the cell viability and the appearance of bone microarchitecture.


Adjuvant chemotherapy is recommended in high-risk stage II–III colorectal cancer (CC). We examine the effect of daily wheatgrass juice (WGJ) intake in addition to chemotherapy on immune parameters, including IL-6, IL-8, IL-10, IL-12, and white blood cells (WBCs) among CC patients. In a controlled prospective trial, 100 stage II–III CC patients were enrolled. According to patient preference, they were divided into two subgroups, control group and intervention group, 50 patients each, all of whom received the same standard postoperative adjuvant chemotherapy, plus consumption of 60 cc WGJ daily in the intervention group. Blood samples were collected at baseline (T0) and upon treatment termination, 5–6 months later (T1). Cytokine concentrations were assessed using ELISA kits. Anti-inflammatory cytokine IL-10 concentrations were significantly higher in the WGJ group than in the control group at T1. The decline in WBC counts between T0 and T1 was significantly lower in the WGJ group. No significant differences were observed in IL-6, IL-8, and IL-12 concentrations between the study groups. The higher levels of IL-10 and the attenuating of WBC decline during chemotherapy may constitute preliminary evidence of the beneficial effects of WGJ on immune parameters, when given as a supplement to standard care. In light of these preliminary results, WGJ supports immunological parameters during adjuvant chemotherapy. Nevertheless, future studies are needed in order to translate those results to clinical recommendations for cancer survivors. Colorectal cancer (CC) is the third most common type of cancer, accounting for 10% of all cancer cases. It is the second most prevalent type in terms of the number of individuals living with cancer five years after diagnosis, worldwide . The higCytokines are a diverse group of intracellular messengers. They orchestrate and regulate the human immune and inflammatory responses and maintain a proper balance among the various immune cell types ,4. Some Leukopenia, a low white blood cell (WBC) count, is commonly caused by chemotherapy . In partA wide range of health benefits have been attributed to wheatgrass, the young grass of the common wheat plant, Triticum aestivum. Its components include chlorophyll, flavonoids, and a number of vitamins , minerals, amino acids, and vital enzymes ,15. FormLaboratory in vitro studies, mostly using the fermented wheat germ extract, have demonstrated anticancer potential: antiproliferative effects were observed in different cancer cell lines ,17,18. FOverall, possible associations between WGJ consumption and improved immune and inflammatory measures may imply beneficial effects of wheatgrass on the disease course, although there is a lack of established empirical knowledge concerning the subject. Therefore, the current study aimed to examine the possible effects of daily WGJ intake on immune measures: pro- and anti-inflammatory cytokines IL-6, IL-8, IL-10, and IL-12 and WBC change (Δ) during chemotherapy.Baseline demographics, disease stage, chemotherapy regimen, background diseases, and anticoagulants drugs were balanced across the study population. Consequently, there were no significant differences between the two groups regarding the background variables . Patientn = 4), a few days of pause in WGJ consumption were reported due to increased nausea, cold sensitivity, etc., following the chemotherapy infusion. However, after several days’ break, the intake was resumed. In two other cases, a break in consumption for several days due to a temporary departure from the country and the resumption of intake upon return was reported.As part of the follow-up during the study, participants were asked regarding coping with chemotherapy, how well they were handling daily WGJ consumption, and whether any difficulties had arisen. Thus, follow-up of patients in the WGJ intervention group was maintained throughout the study, allowing for assessing the persistence of juice consumption and for responding to questions. Patients demonstrated good adherence to daily juice intake (60 cc) during the study period. In general, patients viewed WGJ consumption as an integral part of treatment, along with the medication routine. Hence, consumption was usually continuous during the treatment period . In a few cases (p < 0.05). Similarly, there was a trend (p = 0.07) for a greater mean difference of neutrophil counts in the control group. Hence, the decrease in total WBC and neutrophil counts during chemotherapy was less in the intervention group, with similar mean differences of lymphocyte and monocyte counts in both groups.Similar mean concentrations of cytokines IL-6, IL-8, and IL-12 at T0 and T1 were found in both study groups, with no statistically significant differences . In contp = 0.07) were observed for statistically significant interactions of time with the group variable in both models that predicted total WBC and neutrophil counts, indicating a milder decline in these counts during chemotherapy in the WGJ intervention group. These interactions reveal that the change in WBC counts from baseline throughout chemotherapy and the change in neutrophils in particular were different between the study groups. In addition, the group variable had a significant positive effect on lymphocyte and monocyte counts: the WGJ intervention group was associated with higher counts of lymphocytes and monocytes during chemotherapy.p = 0.47). Grade ≥II nausea and vomiting was less reported in the WGJ group; three patients (6%) compared to eight (16%) in the control arm (0.12). The number of hospitalizations during the treatment period was reported for 25% of the patients, without significant difference of occurrence between the control and WGJ groups (22% in the WGJ and 28% in the control). Two patients died during the treatment period due to intolerable toxicity (one in each group). Median follow-up time was 22 months for both groups.The study was not planned to show differences in clinical outcomes. Nevertheless, the main outcomes are reported . Diarrhep = 0.24). Eleven patients had died at the time of analysis, three in the WGJ group (one due to toxicity and two due to metastatic disease) and eight in the control group .With a median follow-up of 24 months, the number of patients with recurrent disease was 23, with 14 in the WGJ group compared with 9 in the control group (p = 0.14). There was no statistically significant difference in all clinical parameters. Although the OS result favored wheatgrass supplementation, this may also have partly related to a higher percentage of control patients with Stage III CC (94% versus 80%) at baseline.Overall survival (OS) in the 1st, 2nd, and 3rd year was 96%, 87%, and 84%, respectively, in the control group and 98%, 96%, and 96%, respectively, in the WGJ group  have also demonstrated higher counts of lymphocytes of different types in the intervention group among cancer patients ,55,58,59The present study also indicates higher counts of monocytes (phagocytes cells that protect the body against infection) in the intervention group. Monocyte counts increased during chemotherapy in both groups, with a steeper increase in the intervention group. IL-10, which regulates and restrains the inflammatory process, is usually secreted by immune cells, particularly monocytes . TherefoA possible elucidation for the higher monocyte and lymphocyte counts, milder decline in WBC, and a higher concentration of IL-10 observed in the WGJ intervention group emerges from a study examining the effect of oligosaccharides isolated from wheatgrass on human peripheral blood mononuclear cells . AccordiIn summary, differences were observed in all WBC subpopulations examined between the two study groups. Both current and prior studies suggest that wheatgrass has the potential to serve as a supportive therapy during chemotherapy, with a protective effect from leucopenia and the adverse effects associated with it.As mentioned earlier, the study was not planned to show differences in clinical outcomes. However, several chemotherapy side-effects were reported. Numerically, fewer patients in the WGJ group had nausea and vomiting or severe diarrhea. Nevertheless, with respect to hospitalizations during treatment, no significant differences were found between the groups. The chemotherapy protocol based on 5-FU and oxaliplatin used for colon cancer usually does not cause severe neutropenia. Although relief from hematologic side effects (decreased WBC counts) was demonstrated in the present study, it was not translated to clinical outcomes.p = 0.14). Based on the difference in survival after 24 months of follow-up seen in the current study, the number of patients needed to be treated is 268 in order to reach significance. This study is under-powered to reach any conclusion regarding survival benefits of WGJ during the chemotherapy period. It may also logically hypothesize that the most significant effect of WGJ on disease course and survival is during juice intake, and not necessarily a considerable period of time after cessation of consumption. Therefore, in order to answer the question of survival advantage, a larger study with a longer period of WGJ consumption is needed.The results of the current study may indicate a survival benefit for patients in the WGJ group; this is in line with the beneficial effect of WGJ on immune functions demonstrated in this study. However, this benefit was not significant (n = 5). The selection of suitable candidates for the study was conducted through the hospital’s computerized system. Letters were sent to suitable candidates to inform them of research being conducted in the Division of Oncology at RHCC. A week later, the patients were telephoned to suggest participation in the study, as well as to provide information and explanation regarding the study and to answer questions. Response rate was 70.5%.One hundred patients were recruited consecutively between 6 March 2014 and 11 July 2017. All procedures performed in studies involving human participants were in accordance with the ethical standards of the institutional and/or national research committee and with the 1964 Helsinki declaration and its later amendments or comparable ethical standards. All participants signed an informed consent form for inclusion prior to their participation. The study was approved on 9 December 2013, by the Institutional Review Board of Rambam Health Care Campus (RHCC) in Haifa, Israel  (NIH number NCT01991080). The present study is part of a larger research project conducted with regard to WGJ consumption in relation to extracellular vesicles (EVs) and psychological variables. The inclusion criteria were as follows: patients aged 18+, diagnosed with CC stages II–III toward adjuvant chemotherapy following curative surgery. Exclusion criteria were: patients with an infectious disease (hepatitis B), or with difficulty understanding the Hebrew language. However, the number of patients discharged due to these causes was low . The period of WGJ intake was identical to the chemotherapy period for the relevant group. Consistent follow-up with all patients was maintained throughout the study period (control and intervention groups). Monitoring and communication with participants were conducted equally for both groups. The follow-up was usually maintained via a personal frontal conversation when patients arrived at RHCC to receive chemotherapy treatment or to visit the attending physician, once every two to three weeks. If this was not possible, a telephone call was made instead. During these follow-up conversations, it was possible to monitor patients’ well-being, coping with chemotherapy, difficulties that arose, and adherence to WGJ intake.In the current study, we used pure natural WGJ without any processing or additives, supplied by a single agriculturist. The grass was grown on unified compost, consisting entirely of organic manure. Wheat seeds were germinated on trays that contain the compost throughout the year. There was an automatic constant irrigation during the growth. Harvest took place whenever the wheat sprouts reached a length of about 15–20 cm . The content of nutritional components of the wheatgrass grown in this method does not change throughout the year . Squeezing was always cold press-done by a special slow rpm juicer. As a result, the grass, as well as the juice, stays cool throughout the process, and nutritional values are preserved. Moreover, squeezing was carried out immediately before freezing in order to maintain these nutritional values. Each patient in the intervention group received a monthly supply of frozen WGJ, divided into daily rations of 60 cc, during the chemotherapy treatment period (5–6 months). The juice was kept in a freezer, and the participants were asked to drink one dose every morning on an empty stomach.Blood samples were obtained from each patient before the first chemotherapy cycle  and after completion of the chemotherapy treatment 5–6 months later . Briefly, 15 mL of peripheral venous blood were drawn from study participants into ethylenediaminetetraacetic acid (EDTA) tubes; plasma was obtained after two centrifugations  within an hour of collection and frozen at −80 °C. Samples were used to assess pro- and anti-inflammatory cytokine concentrations using appropriate kits .https://www.biolegend.com/Files/Images/media_assets/pro_detail/datasheets/430504_V02.pdf).For cytokine quantification, the following kits were used: Human IL-6 ELISA MAX Deluxe, BioLegend, San Diego, CA, USA. cat # 430505; Human IL-8 ELISA MAX Deluxe, BioLegend, San Diego CA, USA. cat # 431505; Human IL-10 ELISA MAX Deluxe, BioLegend, San Diego, CA, USA. cat # 430605; Human IL-12 p70 ELISA MAX Deluxe, BioLegend, San Diego, CA, USA. cat # 431705  was performed according to the manufacturer’s instructions. Briefly, a day prior to the experiment, the plates were coated with a primary (Capture) antibody  and incubated overnight at 4 °C. On the experiment day, washing was performed four times , and the plates were blocked. Standard and samples (100 µL) were added in duplicates to the wells containing capture antibodies and incubated for two hours with gentle shaking (the samples were not diluted). Biotinylated detection antibody  was then added to the wells. After 60 min of incubation, avidin–HRP (horseradish peroxidase) complex was added. Thirty minutes later, the substrate (tetramethylbenzidine substrate solution) was added, and the plates were incubated in the dark. At this point, a blue dye was formed, which was proportional to the concentrations of each of the cytokines in the blood samples. The reaction was stopped by adding 100 μL of acid solution to each well, and the plates were read at 450 nm.Clinical data at T0 and WBC data at T0 and T1 were collected from the patient’s medical record, as these are routine tests for patients receiving this chemotherapy regimen. Apart from total WBC, subpopulations of neutrophils, lymphocytes, and monocytes were also collected and studied. Demographic data were collected using a self-report demographic questionnaire.t test were used to determine the difference between patients’ demographic and clinical characteristics in both groups. Differences between the study groups regarding the immune variables at T0 and T1 were calculated using the independent t tests. In order to test these differences while adjusting for background variables, a linear mixed effect regression (LMM) was used.Statistical analyses were performed using SPSS (version 23) software for Windows. The chi square test and the independent The main advantage of LMM lies in the fact that the analysis includes the data of all study participants, including patients who had not completed chemotherapy (according to intention to treat approach). From the clinical and demographic variables, only those variables that were correlated significantly with the outcome variables were entered into the LMM models and adjusted for. They were entered to the models as covariates together with the group variable, the time variable, and the interactions between time and group. The group variable examined whether the intervention had an effect on the immune variables, compared with the control group; the time variable examined whether time had an effect on the immune variables; and the interaction between time and group tested whether the change over time in the immune variables differed between the two groups. The background variables were added to the models in order to adjust for their effect on the relationships between group and time variables and the immune variables.Background variables that were significantly associated with the outcome variables (immune measures) were: education (for IL-6); gender, education, background diseases (for IL-8); gender (for IL-10); education, smoking (for IL-12); smoking (for WBC); employment (for lymphocytes); gender, disease stage (for monocytes). No significant associations between neutrophils and background variables were found.p Values of 0.05 were considered statistically significant. However, a statistical significance level of p < 0.1 was presented as well. This is due to the difficulty of reaching statistical significance when the dependent variable is biological, similar to previous studies [ studies ,66.The estimated sample size was calculated according to Cohen’s equation; in a regression model with six independent variables, in order to achieve a medium effect size, at a significance level of 0.05, and a statistical power of 0.80, a sample of 100 participants was required .The main limitation of this study is the non-random allocation into intervention and control groups, based on self-selection. This could result in patient-related bias (volunteer bias). However, no statistically significant differences were found in baseline demographic and clinical characteristics between the study groups. Moreover, the baseline characteristics that were significantly associated with the outcome variables (immune measures) were added to the LMM regression models in order to adjust for their effect on the relationships between WGJ intervention, time, and the immune outcome variables. Further, the same characteristics that motivated patients in the study sample to choose WGJ consumption most likely resemble the characteristics that would motivate patients in the general population to consume WGJ . It is therefore plausible to assume that the findings of the current study would apply to the general population of CC patients, despite self-selection. In addition, although the study was controlled, a placebo effect was not tested.This is a preliminary study devised to examine the possible effects of WGJ on the immune system. Our findings in relation to the anti-inflammatory cytokine IL-10, as well as WBC counts during chemotherapy, constitute preliminary evidence highlighting the beneficial effects of WGJ on immune parameters, when given as a supplement to standard care. There is preliminary evidence, both from the current and previous studies, that wheatgrass has the potential to mitigate a number of different chemotherapy-induced damages. Moreover, wheatgrass constitutes no adverse effects comparable to chemical drugs. Hence, it has a potential to be used as a cheap, non-toxic supplement to standard chemotherapy. The present study, therefore, contributes to the creation of evidence-based recommendations for the use of WGJ among CC patients during chemotherapy.


Post hoc analysis showed a significant reduction in stiffness during hypergravity (p < 0.001) and an increase in stiffness during microgravity (p < 0.001). Activity in all muscles significantly increased during hypergravity (p’s < 0.001). During microgravity, the multifidi (p < 0.002) and transversus abdominis (p < 0.001) increased significantly in muscle activity while no significant difference was found for the psoas (p = 0.850) and erector spinae muscles (p = 0.813). Lumbar curvature flattened in hypergravity as well as microgravity, albeit in different ways: during hypergravity, the distance to the skin decreased for the upper (p = 0.016) and the lower sensor (p = 0.036). During microgravity, the upper sensor showed a significant increase (p = 0.016), and the lower showed a decrease (p = 0.005) in distance. This study emphasizes the role of spinal motor control adaptations in changing gravity conditions. Both hypergravity and microgravity lead to changes in spinal motor control. The decrease in spinal stiffness during hypergravity is interpreted as a shift of the axial load from the spine to the pelvis and thoracic cage. In microgravity, activity of the multifidi and of the psoas muscles seems to ensure the integrity of the spine. Swiss (BASEC-NR: 2018-00051)/French “EST-III” (Nr-ID-RCB: 2018-A011294-51/Nr-CPP: 18.06.09).The objective of this study was to determine the response of the lumbar spinal motor control in different gravitational conditions. This was accomplished by measuring indicators of lumbar motor control, specifically lumbar spinal stiffness, activity of lumbar extensor and flexor muscles and lumbar curvature, in hypergravity and microgravity during parabolic flights. Three female and five male subjects participated in this study. The mean age was 35.5 years (standard deviation: 8.5 years). Spinal stiffness of the L3 vertebra was measured using impulse response; activity of the erector spinae, multifidi, transversus abdominis, and psoas muscles was recorded using surface electromyography; and lumbar curvature was measured using distance sensors mounted on the back-plate of a full-body harness. An effect of gravity condition on spinal stiffness, activity of all muscles assessed and lumbar curvature ( Astronauts who are exposed to microgravity often report lower back pain (LBP) that can significantly affect their ability to work . Total uTherefore, the objective of the current study was to determine the response of the spinal motor control to MG and HG conditions. This was accomplished by testing the hypotheses that lumbar spinal stiffness, lumbar extensor and flexor muscles, and lumbar curvature change with different gravity conditions during parabolic flight. Spinal stiffness is the result of the integration of passive, active and neural motor control subsystems and thereby serves as a proxy measure of spinal motor control . The curEight healthy participants  participated in this study. Participants passed the required aviation medical screening during which neural or musculoskeletal disorders were excluded  and provA single-group repeated-measures design was used to measure changes in spinal motor control evoked by HG and MG. HG and MG were induced using parabolic flight. Parabolic flights offer a sequence of consecutive gravity conditions including 1 g Earth gravity (EG), 1.8 g HG, and μg MG, with small variations of the exact gravitational forces during the different gravity conditions. The course of one parabola started with a horizontal and level flight with normal Earth gravity (EG), followed by a steep climb flight that induced 20 s of HG (1.8 × g). Next, the airplane pushed over the top to begin 22 s of MG within the parabola. Subsequently, a second HG phase followed to return to normal flight level. Measurements were executed in upright posture. Before flight (30 min), participants were administered scopolamine  to prevent motion sickness . The admSpinal stiffness for the purpose of this study was defined as the resistance to deformation of the spinal system, which includes passive rigid structures bones), passive deformable structures , and active structures (muscles) ones, pas. To measOne way to describe the muscular balance of the human spine is to differentiate between local and global muscle systems. The local system consists of muscles with insertion and/or origin at the lumbar vertebrae, the global system of muscles with origin at the pelvis and insertion at the thorax . Here, mLumbar curvature was assessed by two sensors mounted on the aluminum back-plate of the full-body harness. The ultrasonic distance sensors  were integrated into the backplate at + 2 cm rostrally (upper sensor) and −2 cm caudally (lower sensor) to the stiffness device , 2. ThesPer participant, a sequence of 15 parabolas was performed. The first and second parabolas were conducted to familiarize the participants with different gravity conditions and to minimize anxiety. During these first parabolas, the measurement operator secured the participants; therefore, no stiffness measurements were conducted. The final parabola was likewise discarded, as participants were fatigued and already preparing to exit the measurement apparatus. This resulted in a dataset consisting of 12 parabolas per participant. Because respiration can affect spinal stiffness measurements, the participants were instructed to hold their breath at the end of a normal exhalation for the measurement . Tethersparabola = 12). Thereafter, the shortest duration of all detected steady states in all participants was determined, which was used as the data extraction window (tSteadyState = 11.4 s). All EMG channels were segmented into the gravitational steady states, and signals of tSteadyState duration were extracted for each participant, parabola, and muscle. Extraction algorithmically attempted to maximize the distance of this window from any state change while minimizing spectral and raw signal variance. Each sequence was visually verified before calculating the root mean square (RMS). In the case of visible aberrant spectral content or raw signal artifacts in the raw signal, windows were manually shifted to reduce these. For each parabola, RMS of the first EG, MG, and HG state were normalized to the average RMS of EG1 and EG2 states. The resultant ratio was used for further analysis. The normalization to average EG was performed within each parabola individually, as there was large inter-parabola variance in the RMS extracted for the EG states. Within-parabola, the two EG states demonstrated good agreement. Finally, for each muscle and participant, the relative RMS ratio was averaged over all twelve parabolas and for left and right sides. This resulted in a single average RMS value for each muscle and participant. This method was selected in order to avoid averaging raw EMG signals from all parabolas – an approach that could result in contamination of the RMS through microvariations of the gravitational steady state (± 0.1 g). An example of raw data is found in Muscle activity data was processed in MATLAB . Using the mean left and right vertical accelerometer traces of the erector spinae, gravitational steady states  were identified for each parabola  of the t-tests family . The data was transferred, stored, and analyzed using the IBM SPSS 25 statistical software package .We used the Shapiro–Wilk test to test data for normal distribution. In case of a non-normal data distribution differences across all three gravity conditions were analyzed using the Friedman Test. For  effects . Power w G∗power  for the 2(2) = 196.395; p < 0.001]. Results are shown in Post hoc analysis showed significant reduction in stiffness during HG (p < 0.001) and increase in stiffness during MG (p < 0.001).Shapiro–Wilk testing indicated that the data were not normally distributed. The Friedman test showed a statistically significant influence of gravity conditions on spinal stiffness ; multifidi , transversus abdominis , and psoas . Post hoc analysis showed significant increase in muscle activity in all muscles during HG (p < 0.001). During MG the multifidi (p < 0.002) and transversus abdominis muscle activity (p < 0.001) increased significantly. In contrast, no significant change of the psoas  and erector spinae (p = 0.813) muscle activity was found during MG.There was a significant effect of gravity condition on all muscles assessed: erector spinae . Post hoc analysis showed a significant decrease in distance in the upper (p = 0.016) and lower (p = 0.036) sensors during HG. During MG, the upper sensor showed a significant increase (p = 0.016), and the lower showed a decrease (p = 0.005) in distance. A schematic of the changed distances across conditions between sensors and lumbar back are shown in post hoc analysis, effect sizes, and power calculations are shown in Gravity had significantly effect on both lumbar distances [upper sensor: χThis study investigated the role of spinal motor control strategies maintaining lumbar spine integrity during changing gravity conditions. Spinal stiffness increased during HG and decreased during MG, confirming the differential changes in spinal stiffness during HG and MG previously observed in a single-case study . Spinal in vivo assessment of spinal stiffness as performed here represents the net effect of all subsystems, it appears that the effects of the increased activation of the global muscle system dominated over the effects of the local muscle system and the flattening of the lumbar spine. Such dominance of the global system, which supports the global mechanical model of the spinal system by Concurrently with the decrease in lumbar spinal stiffness during HG, an increase in lumbar muscle activity and a flattening of the lumbar curvature occurred. Flattening of the lumbar curvature during HG corroborates findings of a study in which participants were carrying heavy backpacks of up to 60% of their body weights . Both, fMG led to an increase in lumbar spinal stiffness, an increase in multifidi muscle activity, a decrease in transversus abdominis muscle activity, and a flattening of the lumbar curvature. In contrast to the HG condition, no change in muscle activity in the erector spinae and the psoas was detected.As discussed in “Hypergravity.” the decrease of spinal stiffness during HG can be explained by the local and global mechanical model of the spinal system. Similarly, the increase in spinal stiffness during MG can be explained by the observed activity increase of a local stabilizer, the multifidus muscle, and the observed activity decrease of a global stabilizer, the transversus abdominis muscle. In contrast, no change of the activity of the psoas and the erector spinae muscles was detected during MG. Therefore, the reaction of the muscles measured during the present study fits only partially to the local and global mechanical model of the spinal system. During long exposure to MG, e.g., during space missions, the erector spinae shows a similar reduction in activity and muscle mass degeneration as other spinal muscles . Thus, iSimilar to HG, a small flattening of the lumbar curvature occurred during MG, which might have contributed to the observed increase in spinal stiffness. However, it occurs that the shape of the flattening differs between HG and MG. During HG, both sensors showed a decrease in distance, which is expected by the load shift away from the spine by the dominance of the global stabilizers. In contrast during MG, the lower sensor showed a decrease whereas the upper showed an increase in distance. This is expected with the largely reduced gravitational weight that leads to an overall flattening of all spinal curvatures . A modelIn this study, the spinal curvature was measured with distance sensors at two locations of the lumbar spine. Because only two points were measured, these measurements provide a limited assessment of the spinal curvature. Although significant changes were observed, more measurement sites should be used in future studies to obtain more detailed information about curvature changes. Also, it might be argued that the psoas EMG signal is contaminated with crosstalk from the sartorius muscle, however, this has been previously quantified as negligible with appropriate electrode placement .This study emphasizes the role of adaptations of spinal motor control under changing gravity conditions. Both HG and MG lead to changes in spinal motor control. During HG, the axial load appears to shift from the spine to the pelvis and thoracic cage, whereas in MG, ongoing activity of the erector spinae muscle and of the psoas muscle seems to ensure the integrity of the spine. Furthermore, when astronauts return to earth, their motor control must be re-adapted back to EG. This suggests that exercises for spinal motor skills should also be performed under different axial load situations as they occur in everyday life to reduce possible risk of injury to the spine.The raw data supporting the conclusions of this article will be made available by the authors, without undue reservation, to any qualified researcher.The studies involving human participants were reviewed and approved by the Ethics Committee of the Canton of Zurich, Switzerland (BASEC-NR: 2018-00051), and the French Comite de Protection des Personnes EST-III (Nr-ID-RCB: 2018-A011294-51/Nr-CPP: 18.06.09). The patients/participants provided their written informed consent to participate in this study.JS and PS developed the research questions and the design. JS and AL conducted the data acquisition. JS and CE carried out the analysis and interpretation of the results. JS produced an early version of the manuscript. JS, AL, MM, CE, OU, and PS revised the manuscript to bring it to its current version. All authors contributed to the article and approved the submitted version.The authors declare that the research was conducted in the absence of any commercial or financial relationships that could be construed as a potential conflict of interest.


Valeriana officinalis  in treatment of some types of headache, the effect of valerian root was studied in patients with tension-type headache.Tension-type headache is the most frequent type of headache. Considering the effectiveness of The current study is a double-blind randomized clinical trial that was conducted in Shams Hospital of Mashhad University of Medical Sciences, Mashhad, Iran, from January to June 2018. We included 88 participants with tension-type headache and randomly assigned them to intervention and control group by block randomization in a 1:1 ratio. The intervention group received Sedamin® capsule  while the placebo group received 500 mg of breadcrumbs both given as two capsules daily for a month -after dinner. The headache impact on activity of daily livings performance, headache disability, and headache severity were measured using questionnaires in baseline and one month after intervention in both groups.The average age (±SD) of the participants was 34.9 (±8.7) years old. After one month, the impact of headache on daily livings performance, significantly reduced in intervention group (mean=51.2) versus the placebo (mean=57.0), (p<0.001). There was a significant reduction in disability in intervention group (mean=22.9) compared to the placebo (mean=27.4) (p<0.001) and the severity score showed significant reductions in intervention group (mean=3.5) versus the placebo group (mean=5.1) (p<0.001).The present trial showed that valerian capsule could reduce the headache impact on daily livings performance, disability and severity of tension-type headache. Headache is one of the most common medical complaints and disorders, worldwide . Persian medicine is a kind of traditional medicine and its most important approach is finding the fundamental causes of diseases on one side and studying the best treatment strategy on the other side. For example, in Persian medicine, impairment of the gastric function and lower esophageal sphincter (LES), and brain dysfunction, are among the important causes of headache. It seems that plants that strengthen the LES and improve stomach function, as well as the brain tonics can play a role in reducing headache. Since valerian can strengthen the LES, it can be a good choice for the treatment of headache because of its tonic effects on gastric function and LES.According to the WHO, “Valeriana officinalis L.  is a plant from the Caprifoliaceae family, which is native to different temperate climates of Europe, Asia and America  that was done in accordance with the 2010 Consort Guidelines and conducted in the neurology clinic of Shams Hospital of Mashhad University of Medical Sciences, Mashhad, Iran, from January to June 2018. This clinical trial was approved by the ethics committee of Iran University of Medical Sciences (IUMS), Tehran, Iran (No. IR.IUMS.FMD.REC1396.9321309012) and registered at Iranian Registry of Clinical Trials (IRCT) under the registration code IRCT20171203037738N1. We included 88 patients suffering from tension-headache in this RCT. The patients were randomly allocated in two groups equally using the blocked randomization (four blocks) method. The numbers were written on cards inside sealed, opaque envelopes and were kept by a secretary who was not aware of A&B codes and A&B drug box.First, all patients who referred to the clinic were evaluated to select those who meet the inclusion criteria. Second, after clinical evaluations done by a neurologist and checking the inclusion criteria, they were introduced to a trained secretary. Next, the secretary allocated a random card A or B to them. In the final stage, all the included participants were referred to the investigator and they filled a self-administered questionnaire and received intervention based on the card received. In addition, considering the double-blind nature of the study, neither the researcher nor the patients knew which card they had got. All participants signed a written informed consent. The duration of the intervention was one month and researcher was on call (24 hr) during the whole study period. Compliance with treatment during the study was assessed and further reinforced by the investigator in weekly direct contact with all participants. Patients were visited weekly by a neurologist during the study period. If a patient felt a need to take a daily painkiller due to severe headache, then, that participant would be excluded during the intervention. After one month, both groups were invited to the clinic and were asked to fill the same questionnaire again. During the study, participants continued their previous medications.Drug and placebo preparation V.officinalis known as medicinal part of plant were considered in this study  were purchased from Goldaru pharmaceutical company, Isfahan, Iran. Each capsule contained 530 mg of valerian root extraction. Its health code was 1228022753 and medication barcode was 6260232370041. The valerian and placebo capsules were similar in size, appearance, and color and marked with code A and B. Sixty capsules were given to the participants (to take two capsules daily after dinner) for one month.The dried roots of ParticipantsAll the patients with headache who came to the clinic, were evaluated by the neurologist on the basis of the International Classification of Headache Disorders criteria (ICHD-III)   analysis, Mann–Whitney U test, independent samples t-tests, paired t-test, Wilcox, and chi-square tests were used for bivariate comparisons between groups. Statistical analysis was done using SPSS 16 software . Baseline measurementsFrom January to June 2018, 110 patients were recruited for the present trial. Out of 110 participants, 22 did not satisfy the inclusion criteria and were therefore excluded from the study. The reasons for exclusion were: not eligible based on inclusion criteria (n=11), declined to participate (n=6), loss to follow-up (n=3), history of hypertension (n=1) and pregnancy (n=1). Finally, 88 persons were included in the study and randomly allocated to treatment (n=44 participants) and control group (n=44) . The meaMain findingsThe mean difference and 95% confidence interval (CI 95%) of HIT-6, HDI and VAS questionnaires in baseline and one month after intervention are shown in Adverse eventsNo drug complications were reported in the treatment and control groups during the study period. However, one participant in valerian group was diagnosed with dizziness at the beginning of the treatment. Therefore, we divided the dosage of intervention to one capsule at night and another in the morning.In this study, the effect of valerian on tension-type headache was evaluated and compared with placebo in a randomized, double-blind, placebo-controlled study. Findings showed that valerian capsule has a significant effect on the severity and disability of tension-type headaches compared to placebo. This finding was obtained by three HIT-6, HDI, and VAS questionnaires which measured the endpoints before and after the intervention.The pharmacological effects of valerian are attributed to the constituents of volatile oils, monoterpenes, valepotriates and sesquiterpenes  . Some of these constituents were reported to act directly on the brain while valerenic acid inhibits enzyme induced breakdown of γ-amino butyric acid (GABA) in the brain which finally results in sedation  on migraine was evaluated for eight weeks. The finding showed that in this group, eight weeks of activity significantly reduced the number of headache days, headache severity, anxiety and depression symptoms which refer to valerian's effect on the migraine headache. (Mirzaee et al., 2015According to Persian medicine, valerian plays a role in relieving headaches by improving the brain function. Several studies showed that valerian exerts a significant positive effect on treatment of problems such as insomnia, bad sleep, anxiety, and depression. There are some studies about the analgesic effect of valerian on migraine and dysmenorrhea. In a randomized clinical trial study, the effect of Similar studies in children and adolescents with different doses of valerian could be designed. Phytochemical study for indicating the active compounds of valerian in tension-type headache is recommended. In the present study, we showed that valerian capsule could reduce the headache impact on performance of daily livings, severity, and disability of a tension-type headache. So, it can be an effective treatment that could be recommended as a therapy to the patients who suffer from this type of headache along with other common drugs.


Pseudomonas aeruginosa is a public health problem causing both community and hospital-acquired infections, and thus the development of new therapies for these infections is critical. The objective of this study was to analyze in vitro the activity of pentamidine as adjuvant in combinations to antibiotics against seven clinical P. aeruginosa strains. The Minimum Inhibitory Concentration (MIC) was determined following standard protocols, and the results were interpreted according to the European Committee on Antimicrobial Susceptibility Testing (EUCAST) breakpoints; however, the gentamicin activity was interpreted according to the Clinical and Laboratory Standards Institute (CLSI) recommendations. The bactericidal in vitro activity was studied at 1×MIC concentrations by time–kill curves, and also performed in three selected strains at 1/2×MIC of pentamidine. All studies were performed in triplicate. The pentamidine MIC range was 400–1600 μg/mL. Four of the strains were MDR, and the other three were resistant to two antibiotic families. The combinations of pentamidine at 1×MIC showed synergistic activity against all the tested strains, except for pentamidine plus colistin. Pentamidine plus imipenem and meropenem were the combinations that showed synergistic activity against the most strains. At 1/2×MIC, pentamidine plus antibiotics were synergistic with all three analyzed strains. In summary, pentamidine in combination with antibiotics showed in vitro synergy against multidrug-resistant P. aeruginosa clinical strains, which suggests its possible use as adjuvant to antibiotics for the therapy of infections from MDR P. aeruginosa.Multidrug-resistant (MDR)  Pseudomonas aeruginosa is an opportunistic pathogen associated with an ever-widening array of life-threatening acute and chronic infections, including diverse respiratory-tract infections , urinary tract infections, bacteremia, and central nervous infections [P. aeruginosa are a major worry as their spread and mortality continue to increase. These infections are majorly attributed to biofilm formation plays an essential role in the attachment of this bacteria on surfaces, and then contributing to multi-drug resistance in this pathogen [fections . Moreovepathogen .Staphylococcus aureus, Klebsiella pneumoniae, Acinetobacter baumannii, P. aeruginosa, and Enterobacter species), with a worldwide impact on hospital infections and an extensive ability to develop resistance to antibacterial drugs. Thus, P. aeruginosa is one of the pathogens included in the critical category of the World Health Organization (WHO)’s priority list of bacterial pathogens for which research and the development of new antibiotics is urgently required [P. aeruginosa was reported by the European Centre for Disease Prevention and Control (ECDC) Epidemiological Report 2016 on healthcare-associated infections, to be the most frequently isolated microorganism in intensive care unit (ICU)-acquired pneumonia and one of the most frequently isolated in blood-stream infections and urinary tract infections [P. aeruginosa is one of the pathogens included in the group “ESKAPE” , resistance to three or more antimicrobial families among piperacillin/tazobactam, ceftazidime, fluoroquinolones, aminoglycosides, and carbapenems in Germany, Spain and Greece are 6.0%, 10.9%, and 28.7%, respectively. A recent study in India observed that the majority of hospital-acquired isolates were MDR, pointing out an 89.5% of analyzed P. aeruginosa isolates sensitive only to colistin [The 2018 ECDC surveillance of antimicrobial resistance in Europe report  detailedP. aeruginosa are associated with a higher morbidity and mortality, and the associated healthcare burden requires an urgent focus on infection prevention, adequate infection management, and the development of novel therapies to treat this infections [Finally, infections caused by multidrug-resistant (MDR) fections ,9.P. aeruginosa has several advantages, such as reducing the time, cost, and risk associated with the development of new antimicrobials [As a new strategy, the repurposing of drugs for the treatment of infections by MDR crobials . The repcrobials ,12. The crobials .A. baumannii [A. baumannii, finding that the type of carbon source and the availability of iron affect the pentamidine activity against A. baumannii [Pentamidine isethionate, an antiprotozoal drug commonly used in the treatment of trypanosomiasis, leishmaniosis, and fungal infections  was receaumannii . Likewisaumannii . The synaumannii  so it coaumannii . Anotheraumannii .P. aeruginosa and the in vitro and in vivo synergistic effect of pentamidine combined with antibiotics against other GNB, the aim of this study was to evaluate the in vitro activity of pentamidine alone and in combination with different antimicrobials to know if it could be used as an adjuvant to antibiotics commonly used to treat MDR P. aeruginosa infections.Considering the worldwide MDR of The MIC/MBC results of each antimicrobial tested for the different clinical strains are shown in Heteroresistance was observed only with ciprofloxacin for the Pa29 isolate, in which the MIC change from 0.5 mg/L to 4 mg/L after 48 h of incubation at 37 °C.10 CFU/mL. Pentamidine alone at concentration equivalent to 1×MIC showed bactericidal effect against six out of the seven strains. As detailed in the table, pentamidine at 1×MIC shows an early bactericidal activity in 4 of the 6 strains from 2 hours of incubation, and with duration of 24 h in two of the strains. As expected, due to its in vitro activity, the bactericidal activity of antibiotics tested was found only against four strains, and heterogeneous: tobramycin, meropenem, and levofloxacin against Pa194, levofloxacin against Pa206, amikacin and colistin against Pa215, and aztreonam and levofloxacin against Pa249. None of the antibiotics nor pentamidine showed bactericidal activity against the Pa223 strain.The bactericidal activity of the drugs alone is shown in The synergistic and bactericidal activities of the combinations of pentamidine with the antibiotics are shown in The activity of pentamidine at sub-MIC concentration (1/2×MIC) in combination with the selected antibiotics is detailed in P. aeruginosa infections, finding synergistic effect as well as bactericidal activity against most of the strains, when combined pentamidine with most of the tested antibiotics, such as aminoglycosides, carbapenems, aztreonam, ceftazidime, and fluoroquinolones, but not with colistin. Additionally, pentamidine at a sub-inhibitory concentration potentiated the in vitro activity of antibiotics with no in vitro activity when studied alone against three selected P. aeruginosa strains.This study evaluate in vitro the use of pentamidine as adjuvant to antibiotics commonly used to treat MDR P. aeruginosa clinical strains. Given that pentamidine is currently used as an antifungal agent [Escherichia coli strain, with MIC values ranging from 100 to >200 mg/L [Pentamidine alone showed a high MIC values’ range, from 400 to 1600 mg/L, against the seven al agent , there a200 mg/L . Also, t200 mg/L ,15,19.A. baumannii and E. coli [When pentamidine was analyzed alone, in time-kill curves at concentrations equivalent to its MIC values, we found that it exhibits bactericidal activity against six of the seven tested strains. This data, is in accordance with the ones in which pentamidine alone at MIC concentrations showed bactericidal activity in vitro against carbapenemase-producing and/or colistin-resistant Enterobacteriaceae , and wit E. coli . Additio E. coli .E. coli strain expressing a truncated lipopolysaccharide variant no enhanced activity of pentamidine when analyzed alone, fact that would not support the hypothesis of pentamidine activity on an intracellular target. Nevertheless, this finding was supported by experiments using only one strain of E. coli. In our study, we have analyzed, using seven strains of P. aeruginosa, if pentamidine enhances the activity of antibiotics to which the strains were resistant. In this sense, our data shows that pentamidine, at concentrations equivalent to its MIC, increased the activity of most antibiotics tested against the MDR clinical P. aeruginosa strains, regardless if their mechanism of action, intracellular or on the bacterial membrane. Therefore, we observed that this activity was enhanced in antibiotics with intracellular mechanisms of action, such as aminoglycosides  and fluoroquinolones (ciprofloxacin and levofloxacin), but it did also inhibit antibiotics that act by inhibiting the synthesis of the cell wall-like carbapenems (imipenem and meropenem) or cephalosporins (ceftazidime). Furthermore, and regardless of the mechanism of action of carbapenems, pentamidine plus imipenem and meropenem were the combinations that showed synergistic activity against more strains, six and five out of the seven strains tested. These data suggest that further studies are necessary to elucidate the mechanisms of action of pentamidine in P. aeruginosa.In the work of Stokes et al.  they staA. baumannii and E. coli [Moreover, our data shows that antibiotics that alone did not have in vitro activity when combined with a sub-inhibitory pentamidine concentration showed bactericidal and synergistic activity against the tested strains. The present results are in accordance to the ones showed by Cebrero-Cangueiro et al., in which synergism was observed with pentamidine plus antibiotics against carbapenemase-producing and/or colistin-resistant Enterobacteriaceae . Stokes  E. coli .P. aeruginosa, suggesting the possible role of pentamidine, as a repurposed drug, for antibacterial use against these difficult to treat MDR pathogens.In summary, our results show in vitro synergy of pentamidine in combination with several families of antibiotics against MDR P. aeruginosa collected from different samples, during the quasi-experimental ecological study CARBAPIRASOA (RAM-PIR-2016-01) were studied: Pa29 from blood; Pa194 from pharyngeal exudate; Pa206 from surgical wound; Pa215 from skin biopsy; Pa249 from venous catheter; and Pa223 and Pa302 from bronchial aspirates. The Pa194 isolate is an IMP-3 producer.Seven MDR  clinical isolates of Pentamidine isethionate and the nine antibiotics  were used as standard laboratory powders from Sigma , except for ciprofloxacin, which was from Fluka . All the powder drugs were resuspended according to laboratory recommendations for in vitro uses.5 CFU/mL. The MIC was evaluated after approximately 18–24 h of incubation at 37 °C and was defined as the minimal concentration of antibiotic that was able to inhibit macroscopic bacterial growth. The MIC results were interpreted according to the European Committee on Antimicrobial Susceptibility Testing (EUCAST) [P. aeruginosa ATCC 27853 was used as quality control strain.To confirm the data from the standard clinical microbiology studies on susceptibility and resistance of the selected strains, the MICs of pentamidine and the nine antibiotics were determined by broth microdilution as recommended by the Clinical and Laboratory Standards Institute (CLSI) , using M(EUCAST)  breakpoi(EUCAST) , and penMinimum bactericidal concentrations (MBCs) were determined by subculturing 100-μL aliquots from inoculated tubes containing antimicrobial concentrations higher than or equal to the MIC of each agent on to antibiotic-free Mueller–Hinton agar. Plates were incubated at 37 °C for 24 h, and colonies were counted. The MBC was defined as the concentration that killed ≥99.9% of the initial inoculum .The heteroresistance assay was also carried out by reading the MIC after 48 h of incubation. Bacterial strains were defined as heteroresistant when they were able to grow at least two dilutions above the previously determined MIC value . All ant5 CFU/mL, and prefixed concentrations of 1×MIC for the antibiotics and pentamidine. Bacterial growth was quantified after 0, 2, 4, 8, and 24 h of incubation at 37 °C by plating 10-fold dilutions on sheep blood agar. To avoid carryover antimicrobial agent interference, the sample was placed on the plate in a single streak down the center, allowed to absorb into the agar until the plate surface appeared dry, and the inoculum was then spread over the plate. The limit of detection was 10 colony-forming unit (CFU)/mL, corresponding to 1 log10 CFU/mL [Bactericidal and synergistic activities of pentamidine, the antibiotics, and pentamidine plus each antibiotic were performed using bacteria in the log-phase according to standard recommendations , by time0 CFU/mL .10 CFU/mL from the initial inoculum; a bacteriostatic effect was established as no change with respect to the initial bacterial concentration during the 24 h, and synergistic effect was defined as a decrease of ≥2 log10 CFU/mL for combined drugs compared with the most active single agent [Bactericidal effect was defined by a decrease of ≥3 logle agent . The expMoreover, and after the results obtained with the time-kill curves studies at concentrations equivalent to 1×MIC of the drugs, we selected three strains  to evaluate if at sub-MIC pentamidine concentration (1/2×MIC) there was still in vitro synergy when combined with selected antibiotics which did not show in vitro activity when tested alone.P. aeruginosa infections, suggesting this combination as a new alternative in the treatment of infections by this pathogen. Further steps, including pharmacokinetic and pharmacodynamic studies and in vivo efficacy evaluation in experimental models of infection, including the dosage and safety, are required to confirm the usefulness of pentamidine in the treatment of infection by MDR P. aeruginosa.In conclusion, these results provide new insights regarding the repurpose of pentamidine as adjuvant to potentiate the activity of antibiotics commonly used to treat MDR


This study aimed to develop a nomogram to predict the overall survival (OS) of these patients.de novo MBC patients diagnosed in 2010–2016 were identified from the Surveillance, Epidemiology, and End Results (SEER) database. They were randomly divided into a training and a validation cohort with a ratio of 2:1. The best subsets of covariates were identified to develop a nomogram predicting OS based on the smallest Akaike Information Criterion (AIC) value in the multivariate Cox models. The discrimination and calibration of the nomogram were evaluated using the Concordance index, the area under the time-dependent receiver operating characteristic curve (AUC) and calibration curves.In this study, we included 7986 patients with de novo MBC. The median follow-up time was 36 months (range: 0–83 months). Five thousand three-hundred twenty four patients were allocated into the training cohort while 2662 were allocated into the validation cohort. In the training cohort, age at diagnosis, race, marital status, differentiation grade, subtype, T stage, bone metastasis, brain metastasis, liver metastasis, lung metastasis, surgery and chemotherapy were selected to create the nomogram estimating the 1-, 3- and 5- year OS based on the smallest AIC value in the multivariate Cox models. The nomogram achieved a Concordance index of 0.723  in the training cohort and 0.719  in the validation cohort. AUC values of the nomogram indicated good specificity and sensitivity in the training and validation cohort. Calibration curves showed a favorable consistency between the predicted and actual survival probabilities.The developed nomogram reliably predicted OS in patients with de novo MBC and presented a favorable discrimination ability. While further validation is needed, this may be a useful tool in clinical practice. Breast cancer is the most common kind of malignancy in females worldwide; it ranks second in contributing to tumor related death in women , 2. ApprBreast cancer tends to be heterogeneous, characterized by diverse histopathologic and molecular features, including age at diagnosis, race, differentiation grade, molecular subtypes, and site of metastasis. These characteristics were previously reported to be associated with survival of de novo MBC patients , 6. ChemA nomogram is a reliable and accurate visualization model utilizing risk factors identified in multivariate analysis; it is widely used for the prediction of survival in oncology , 14. In http://www.seer.cancer.gov/seerstat) was used to identify eligible patients.Data was obtained from the National Cancer Institute’s Surveillance, Epidemiology, and End Results (SEER) program, which consists of 18 population-based cancer registries, for patients diagnosed between 2010 and 2016. SEER is an open-access resource for tumor-based demographic and pathological information, as well as treatment information and patient survival outcomes. SEER*Stat Version 8.3.4  status and sites of distant metastasis in 2010, this was used as the starting point. The inclusion criteria of MBC patients were listed as follows: female, year of diagnosis from 2010 to 2016, older than 18 years old when diagnosed, breast cancer as the first and only malignant tumor diagnosis, histology of infiltrating duct or/and lobular carcinoma, at least one distant site of de novo metastasis. Patients with unknown condition of marital status, race, differentiation grade, T stage, N stage, site of metastasis, or follow-up information were excluded.Demographic variables including age at diagnosis , race  and marital status  or domestic partner). Tumor characteristics included histology , grade , molecular subtype, T stage , N stage , and metastatic site . Therapies included chemotherapy (No/Unknown and Yes), radiation (No/Unknown and Yes), and surgery of the primary tumor ). Estrogen receptor (ER) and progesterone receptor (PR) status were combined as the hormone receptor (HR) status, and the breast cancer molecular subtype was stratified based on joint HR and human epidermal growth factor receptor-2 (HER2) statuses .Patient demographics, tumor characteristics and treatment information were compared using the chi-square test. Overall survival (OS) was defined as the time from breast cancer diagnosis to death from any cause. Patients in the initial cohort were allocated randomly into a training cohort and a validation cohort with a ratio of 2:1. The training cohort was used to develop a nomogram while the validation model was used to validate the model. In the training cohort, the covariates included in the multivariate Cox proportional hazards models were identified by a backward stepwise method based on the smallest Akaike information criterion (AIC) value, which indicated the minimal loss of prognostic information , 16.The nomogram was developed on the basis of independent risk factors and using the “rms” R package. The predictive capacity of the nomogram was assessed using Harrell’s C-index  and the area under the time-dependent receiver operating characteristic curve (AUC), which estimates the probability between the observed and predicted OS. Bootstrapping method with 1000 resamples was utilized to generate the calibration curves for validation of the nomogram in the training cohort and in the validation cohort. The scores of each variable were calculated using the “nomogramEx” package in R. On the basis of the scores of each variable, the total scores for each patient could be calculated.http://www.r-project.org). Statistical significance was assumed at a two-side p value of < 0.05.All analyses were performed with SPSS  and R version 3.6.0  and the validation cohort (N = 2662). The demographic, pathological and treatment information of the two cohorts is listed in Table The included 7986 patients were allocated randomly into the training cohort  and 0.719  in the training and validation cohort, respectively. In the training cohort, the AUC values of the nomogram to predict 1-, 3- and 5-year OS was 0.784 , 0.777  and 0.786 , respectively. In the validation cohort, the AUC values of the nomogram to predict 1-, 3- and 5-year OS was 0.802 , 0.784  and 0.790 , respectively (Fig. The survival of patients with de novo MBC is difficult to predict, because of the lack of prediction models for these patients. In this study, we developed a nomogram to visualize survival of de novo MBC patients identified from the SEER database. This model was validated, and the performance was evaluated. Calibration plots showed an optimal agreement between the observed risks and the estimated risks by the nomogram, indicating the reliability of this model. Discrimination was evaluated by the C-index and AUC values. Both of them indicated good specificity and sensitivity in this nomogram.Several nomograms have been developed to predict the survival of MBC patients –20. C. KMBC is a kind of heterogeneous diseases. Many factors affect the prognosis and therapeutic efficacy of drugs. The molecular subtype is a vital prognostic factor and serves as the cornerstone of treatment , 24, 25.MBC is considered incurable. Systemic therapy remains the mainstay of therapy . Over thThere existed some limitations in this study. Firstly, it was a retrospective study and it was subject to all the inherent biases associated with this type of study design. Furthermore, some prognostic factors were not included in the SEER database, including the number of metastatic lesions, use of endocrine therapy and use of target therapy. Thirdly, the nomogram in our study was validated in the same population and such validation on model performance could be biased. Therefore, the predictive effect of the nomogram needs to be assessed carefully in other cohorts.The developed nomogram reliably predicted OS in patients with de novo MBC and presented a favorable discrimination ability. Using this model, the role of primary tumor surgery and other significant prognostic factors in MBC patients could be estimated. This will guide surgical decision making in clinical practice, although the findings require additional validation.


Ascending thoracic aortic aneurysm (ATAA), seen in adults, is an important cause of morbidity and mortality. In this study, we aimed to evaluate the levels of mean platelet volume (MPV), mean platelet volume-to-platelet count ratio (MPVPCR), mean platelet volume-to-lymphocyte ratio (MPVLR), and red cell distribution width platelet count ratio (RDWPCR) in patients with thoracic aortic aneurysm.105 patients admitted to the emergency department were diagnosed with thoracic aortic aneurysm between January and December 2014, and 100 healthy individuals were involved in this retrospective study. MPV, MPVLR, MPVPCR and RDWPCRs were calculated at the time of admission.P<0.001, P<0.001, respectively), while MPV, MPVPCR, MPVLR and RDWPCR were found to be significantly higher . In the patient group, the high-sensitivity C-reactive protein was significantly higher (P<0.001), and the neutrophil (P=0.062) was also higher. In ROC analysis, MPVPCR had the highest sensitivity (80%) and RDWPCR had the highest specificity (72%).Platelet and lymphocyte levels were found to be significantly lower in the patient group when compared to the healthy group (The results for MPV, MPVPCR, MPVLR and RDWPCR can be evaluated as useful parameters in the emergency clinical approach in the evaluation of inflammatory activity in ATAA patients. More extensive studies are required to address the role of these parameters in determining the severity of the disease. The fact that patients remain asymptomatic until the development of dissection and rupture is especially a cause of mortality and morbidity in patients in ascending thoracic aortic aneurysm (ATAA). These patients are generally diagnosed when evaluating imaging tests performed for other purposes. The risk factors that take part in the formation and development process of the aneurysm are like the risk factors for coronary artery disease. It is known that there are many cytokines, which cause local and systemic effects and medial degenerative changes, in the etiopathogenesis of AA. Therefore, AA is also considered an inflammatory response. In pathogenesis, in addition to lymphocytes and protease activity, the macrophages, which are accepted as the source of pro-inflammatory cytokines such as IL6, have been shown to be involved. The majority of untreated ascending aortic aneurysms progress in a deadly manner, due to rupture or dissection. Therefore, inflammation is important not only in the formation of cardiovascular diseases, but also in the complications that can occur subsequently.Aortic aneurysm (AA) is defined as the permanent expansion of more than 50% of the normal values of the transverse diameter of any segment of the aorta that should be in the range in accordance with the age and body surface of a person. Even though AA can be seen in both thoracic and abdominal aorta, it is estimated that the incidence of thoracic aortic aneurysm (TAA) has increased.Recent studies have shown that mean platelet volume (MPV) can be used as a diagnostic marker for certain inflammatory disorders. MPV is a marker of activated platelets and is associated with different inflammatory conditions. In patients with diabetes mellitus (DM), cardiovascular disease, peripheral artery disease and cerebrovascular disease, increased MPV levels are associated with a low degree of inflammatory status. However, in patients with ulcerative colitis, rheumatoid arthritis and ankylosing spondylitis, high-grade inflammatory diseases, such as Mediterranean fever, are associated with decreased MPV,9.The mean platelet volume-to-platelet count ratio (MPVPCR) has been reported to be predictive of long-term mortality in many diseases, including ischemic cardiovascular diseases, sepsis, and nonalcoholic fatty liver disease.The mean platelet volume-to-lymphocyte ratio (MPVLR) may be an independent indicator for early and late mortality after ST-segment elevation myocardial infarction (STEMI), as well as predicting in-hospital mortality.The red cell distribution width-to-platelet count ratio (RDWPCR) is shown to be predictive of the severity of liver fibrosis in nonalcoholic fatty liver diseaseReducing inflammation in patients with AA can mean a reduction in existing complications. There is still a lack of biomarkers to assess the risk of aneurysm formation, enlargement or rupture. No study evaluating proinflammatory levels of MPV, MPVPCR, MPVLCR and RDWPCR in patients with ascending aortic aneurysms has been determined in the literature. In this study, we aimed to evaluate the levels of MPV, MPVPCR, MPVLR and RDWPCR in patients with thoracic aortic aneurysms.Patients who were admitted to the emergency department (ED) between January 2014 and December 2014, with complaints of chest pain and respiratory distress and identified by ED diagnosis of “aortic aneurysm” and “aortic dissection”, were further analyzed to select only those with thoracic aortic aneurysm and thoracic aortic dissection. In addition, the medical records of hospitalized patients were reviewed for thoracic aortic dissection and aneurysm. In total, 105 patients were included in the retrospective study as a case group. Moreover, a total of 196 healthy individuals selected from the hospital records within the same period were involved in the study as a control group. In the study, patients previously diagnosed with hematological malignancy, chronic obstructive pulmonary disease (COPD), autoimmune liver disease, cirrhosis, metastatic bone marrow infiltration, and acute or chronic inflammatory disease such as physical trauma, tonsillitis, asthma, rheumatoid arthritis and active hepatitis were excluded from the study. Other exclusion criteria were evidence of current or recent treatment (in the past 3 months) with oral or intravenous steroids or other medications that might cause pancytopenia. Clinical, demographic and laboratory data of each patient were obtained and recorded. Then, age and gender distribution were calculated and ATAA, MPV, MPVLR, MPVPCR and RDWPCR ratios were determined. The diagnosis of ascending aortic aneurysm (40 mm≥) was confirmed by examining the patients with contrast-enhanced thorax tomography and then by an independent radiologist. Patients included in the study (study group) were compared with normal individuals with normal ascending aorta diameter, age, gender, hypertension, and DM with similar distribution (control group). This research was approved by the Human Ethics Committee (2016-03/04).th, 75th percentile) as appropriate. Significance test (independent t-test) and Mann-Whitney U test were used for the comparison of research and control groups. Receiver operating characteristic (ROC) curve analysis was performed and optimal cutoff values were determined for inflammation. An alpha error of 5% was accepted.Data were analyzed using SPSS 22.0 . Variables were expressed as mean ± standard deviation (SD) and median (25P>0.05). However, the mean aortic diameter in the case group (42.3±2.7) was significantly higher than the mean in the control group (35.8±3.2)   .P=0.001). Furthermore, when the two groups were compared in terms of MPV values, it was found that the value in the case group (9.7±1.1) was significantly higher than the control group (9.2±0.8) (P<0.001).The median values of platelet and lymphocyte count in the study group were significantly lower  compared to the control group [254.0 (214.0-298.3) and 1.7 (1.4-2.3)], respectively), and this difference was statistically significant  (P=0.013 for RDWPCR only)  in the case group were significantly higher than the mean values of the control group (0.05±0.02 CR only) .In the present study, the optimal cutoff value determined by ROC analysis for MPV was found to be 9.35 , for MPVPCR, this value was 0.04 , for MPVLR, this value was 5.55  and forThis study was designed to determine the inflammatory process in patients with ATAA, as the first research in the literature using measurements such as MPV, MPVPCR, MPVLR and RDWPCR.. Furthermore, it has been shown in previous studies that AA is also an inflammatory process. For example, in histological examinations of TAA patients, in addition to the high levels of inflammatory cells observed in the adventitia layer, T lymphocytes and macrophages appeared in pathogenesis and there were findings supporting the presence of cytokines. However, in accordance with the current research, studies on patients with ATAA and chronic aortic dissection, high levels of hs-CRP and white blood cell levels have been another evidence of the presence of inflammatory status. In inflammatory cases, it is known that certain parameters of the complete blood count diverse in terms of number and quality.In previous studies, it was accepted that AA dilation is na important predictor of coronary artery disease, ischemic stroke, acute myocardial infarction and cardiovascular mortality. However, MPV levels indicate platelet activity more accurately than platelet count. MPV levels have been shown to decrease in diseases such as active ankylosing spondylitis, rheumatoid arthritis, inflammation period of COPD and systemic lupus erythematosus. Similarly, the MPV level was also low in this study . Parallel to these studies, RDWPCR was also high in this study (The high MPVLR values obtained in our study are consistent with the results of a previous study, which indicates that the MPVLR level is an independent indicator for predicting both early and late mortality after STEMI, and in-hospital mortalityis study ., in which MPVPCR was shown to be a strong independent predictor for in-hospital complications and mortality in patients with aortic dissection. Furthermore, as a result of the study findings, cutoff values of MPVLR and RDWPCR were similar to each other in prediction inflammation in patients with ATAA (Our study revealed that the cutoff value of MPVPCR in predicting inflammation in patients with ATAA is comparable to the result of the study by Li et al.ith ATAA .The approach of patients with ATAA should be concerned with monitoring patients after diagnosis, preventing possible complications, controlling blood pressure and any cardiovascular risk factors. Based on the findings of the study, values of MPVPCR, MPVLR and RDWPCR might be proposed as a quick and useful screening tool that can contribute to diagnose and monitor the inflammatory process to help the physician decide whether to request additional imaging studies to confirm or refuse the ATAA diagnosis.The most important of the limitations of this study is that, since it is a retrospective study with a relatively small patient group, the laboratory findings of certain clinical and inflammatory markers, such as interleukin-6, TNF-α etc., were not available. Another limitation is that only ATAA data can be obtained by computed tomography and these results cannot be confirmed by other imaging methods, echocardiography, and magnetic resonance. The last constraint was that the patients included in the study could not be followed in terms of possible complications and consequences.