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In this paper, a new microstrip triplexer is designed to work at 2.5 GHz, 4.4 GHz and 6 GHz for mid-band 5G applications. All channels are flat with three low group delays (GDs) of 0.84 ns, 0.75 ns and 0.49 ns, respectively. Compared to the previously reported works, the proposed triplexer has the minimum group delay. The designed triplexer has 18.2%, 13.7%, 23.6% fractional bandwidths (FBW%) at 2.5 GHz, 4.4 GHz and 6 GHz, respectively. The obtained insertion losses (ILs) are low at all channels. These features are obtained without a noticeable increase in the overall size. A novel and simple resonator is used to design the proposed triplexer, which includes two pairs of coupled lines combined with a shunt stub. A perfect mathematical analysis is performed to find the resonator behavior and the layout optimization. The type of shunt stub is determined mathematically. Also, the smallness or largeness of some important physical dimensions is determined using the proposed mathematical analysis. Finally, the designed triplexer is fabricated and measured, where the measurement results verify the simulations.
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Desenho de Equipamento , Tecnologia sem Fio , Tecnologia sem Fio/instrumentaçãoRESUMO
This research introduces a new designing process and analysis of an innovative Silicon-on-Insulator Metal-Semiconductor Field-Effect (SOI MESFET) structure that demonstrates improved DC and RF characteristics. The design incorporates several modifications to control and reduce the electric field concentration within the channel. These modifications include relocating the transistor channel to sub-regions near the source and drain, adjusting the position of the gate electrode closer to the source, introducing an aluminum layer beneath the channel, and integrating an oxide layer adjacent to the gate. The results show that the AlOx-MESFET configuration exhibits a remarkable increase of 128% in breakdown voltage and 156% in peak power. Furthermore, due to enhanced conductivity and a significant reduction in gate-drain capacitance, there is a notable improvement of 53% in the cut-off frequency and a 28% increase in the maximum oscillation frequency. Additionally, the current gain experiences a boost of 15%. The improved breakdown voltage and peak power make it suitable for applications requiring robust performance under high voltage and power conditions. The increased maximum oscillation frequency and cut-off frequency make it ideal for high-frequency applications where fast signal processing is crucial. Moreover, the enhanced current gain ensures efficient amplification of signals. The introduced SOI MESFET structure with its modifications offers significant improvements in various performance metrics. It provides high oscillation frequency, better breakdown voltage and good cut-off frequency, and current gain compared to the traditional designs. These enhancements make it a highly desirable choice for applications that demand high-frequency and high-power capabilities.
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Desenho de Equipamento , Silício , Silício/química , Semicondutores , Transistores Eletrônicos , Condutividade Elétrica , Fontes de Energia Elétrica , Metais/químicaRESUMO
Background: Natural killer T (NKT) cells are influential immune cells in pregnancy failures, including recurrent spontaneous abortion (RSA). Different approaches are used for these disorders due to their effects on maternal immunomodulation. Aims: In the present study, we compared the effects of two typical immunotherapies (lymphocyte immunotherapy [LIT] and low-dose prednisolone) on CD3+CD56+CD16+ and CD3+CD56+CD8+ cells as two distinct subsets of NKT cells in Women with RSA. Settings and Design: This study was a comparative cohort study conducted from 2021 to 2022. One hundred and five women with RSA were distributed into three treatment groups randomly. Materials and Methods: Fifty women in the group of low-dose prednisolone therapy, fifty women in the LIT group and five women without any treatment as the control group were included in the study. NK and NKT cell subsets were assessed using flow cytometry. Furthermore, the concentration of interferon-gamma (IFN-γ), transforming growth factor-beta (TGF-ß) and interleukin-10 (IL-10) was measured quantitatively using the enzyme-linked immunosorbent assay technique. Statistical Analysis Used: Normality and comparisons between study groups were performed by non-parametric unpaired Mann-Whitney, Kruskal-Wallis rank sum test, and one-way ANOVA. Results: The percentage of CD56dim NK cells was increased after prednisolone therapy, while this population significantly decreased in the LIT group. In contrast to the LIT group, the administration of prednisolone increased CD3+CD8+CD56+ NKT cells (P < 0.0001), which is helpful for pregnancy. The effect of the investigated treatment approaches on the population of peripheral CD3+CD56+CD16+ NKT cells of women with RSA was not adequately significant. The same situation was also observed regarding the serum level of IFN-γ. However, a significant decrease in serum levels of IL-10 and TGF-ß was observed after prednisolone therapy. Conclusion: The lower capability of LIT in changing the population of NKT cells compared to prednisolone therapy may be due to its mechanism of action, which is related to the production of blocking antibodies. These treatment approaches had different effects on NKT cells, indicating that NKT cell population and function can be affected using LIT and prednisolone therapy distinctly. In addition, prednisolone therapy and LIT in women with normal serum levels of IFN-γ have no harmful effects in changing the production of this critical cytokine.
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Microstrip couplers play a crucial role in signal processing and transmission in various applications, including RF and wireless communication, radar systems, and satellites. In this work, a novel microstrip 180° coupler is designed, fabricated and measured. The layout configuration of this coupler is completely new and different from the previously reported Rat-race, branch-line and directional couplers. To obtain the proposed coupler, the meandrous coupled lines are used and analyzed mathematically. To improve the performance of our coupler, an optimization method is used. The designed coupler is very compact with an overall size of 0.014λg2. The obtained values of S21 and S31 are -3.45 dB and -3.75 dB, respectively at the operating frequency, while the fractional bandwidth (FBW) is 56.2%. It operates at fo = 1.61 GHz (suitable for 5G applications) and can suppress harmonics up to 2.17fo. Another advantage of this coupler is its low phase imbalance, while the phase difference between S21 and S31 is 180°± 0.023°. Therefore, our device is a balanced coupler with ±0.3 dB magnitude unbalance at its operating frequency. It is important to note that it is very difficult to find a coupler that has all these advantages at the same time. The proposed 180° coupler is fabricated and measured. The comparison shows that the measurement and simulation results are in good agreement. Therefore, the proposed coupler can be easily used in designing high-performance 5G communication systems.
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Comunicação , Radar , Animais , Ratos , Simulação por Computador , Processamento de Sinais Assistido por ComputadorRESUMO
Global concerns regarding environmental preservation and energy sustainability have emerged due to the various impacts of constantly increasing energy demands and climate changes. With advancements in smart grid, edge computing, and Metaverse-based technologies, it has become apparent that conventional private power networks are insufficient to meet the demanding requirements of industrial applications. The unique capabilities of 5G, such as numerous connections, high reliability, low latency, and large bandwidth, make it an excellent choice for smart grid services. The 5G network industry will heavily rely on the Internet of Things (IoT) to progress, which will act as a catalyst for the development of the future smart grid. This comprehensive platform will not only include communication infrastructure for smart grid edge computing, but also Metaverse platforms. Therefore, optimizing the IoT is crucial to achieve a sustainable edge computing network. This paper presents the design, fabrication, and evaluation of a super-efficient GSM triplexer for 5G-enabled IoT in sustainable smart grid edge computing and the Metaverse. This component is intended to operate at 0.815/1.58/2.65 GHz for 5G applications. The physical layout of our triplexer is new, and it is presented for the first time in this work. The overall size of our triplexer is only 0.007 λg2, which is the smallest compared to the previous works. The proposed triplexer has very low insertion losses of 0.12 dB, 0.09 dB, and 0.42 dB at the first, second, and third channels, respectively. We achieved the minimum insertion losses compared to previous triplexers. Additionally, the common port return losses (RLs) were better than 26 dB at all channels.
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The higher heating value (HHV) is the main property showing the energy amount of biomass samples. Several linear correlations based on either the proximate or the ultimate analysis have already been proposed for predicting biomass HHV. Since the HHV relationship with the proximate and ultimate analyses is not linear, nonlinear models might be a better alternative. Accordingly, this study employed the Elman recurrent neural network (ENN) to anticipate the HHV of different biomass samples from both the ultimate and proximate compositional analyses as the model inputs. The number of hidden neurons and the training algorithm were determined in such a way that the ENN model showed the highest prediction and generalization accuracy. The single hidden layer ENN with only four nodes, trained by the Levenberg-Marquardt algorithm, was identified as the most accurate model. The proposed ENN exhibited reliable prediction and generalization performance for estimating 532 experimental HHVs with a low mean absolute error of 0.67 and a mean square error of 0.96. In addition, the proposed ENN model provides a ground to clearly understand the dependency of the HHV on the fixed carbon, volatile matter, ash, carbon, hydrogen, nitrogen, oxygen, and sulfur content of biomass feedstocks.
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Calefação , Redes Neurais de Computação , Biomassa , Algoritmos , CarbonoRESUMO
The induction of feto-maternal tolerance, fetal non-immunogenicity, and the regulation of mother's immune system are essential variables in a successful pregnancy. Fetal membranes have been used as a source of stem cells and biological components in recent decades. Human amniotic epithelial cells (hAEC) have stem/progenitor characteristics like those found in the amniotic membrane. Based on their immunomodulatory capabilities, recent studies have focused on the experimental and therapeutic applications of hAECs in allograft transplantation, autoimmune disorders, and gynecological problems such as recurrent spontaneous abortion (RSA), recurrent implantation failure (RIF), and premature ovarian failure (POF). This review discusses some of the immunomodulatory features and therapeutic potential of hAECs in preventing infertility, miscarriage, and implantation failure by controlling the maternal immune system.
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Aborto Espontâneo , Infertilidade , Insuficiência Ovariana Primária , Gravidez , Feminino , Humanos , Aborto Espontâneo/terapia , Células Epiteliais , Insuficiência Ovariana Primária/terapiaRESUMO
Cardiovascular diseases (CVD) pose a serious threat to people's health, with extremely high global morbidity, mortality, and disability rates. This study aimed to review the literature that examined the relationship between blood groups and CVD. Many studies have reported that non-O blood groups are associated with an increased risk and severity of coronary artery disease (CAD) and acute coronary syndromes (ACS). Non-O blood groups increase the risk and severity of these conditions by increasing von-Willebrand factor (VWF) and plasma cholesterol levels and inducing endothelial dysfunction and inflammation. They have also been linked with increased coronary artery calcification, coronary lesion complexity, and poor collateral circulation. Blood groups also affect the prognosis of CAD and ACS and can alter the rate of complications and mortality. Several cardiovascular complications have been described for COVID-19, and blood groups can influence their occurrence. No studies have found a significant relationship between the Lewis blood group and CVD. In conclusion, people with non-O blood groups should be vigilantly monitored for cardiovascular risk factors as prevention and proper treatment of these risk factors may mitigate their risk of CVD and adverse cardiovascular events.
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OBJECTIVES: To show that there is relation between neutrophil-to-lymphocyte ratio (NLR) and metabolic syndrome and also, its components. FINDINGS: Metabolic syndrome is associated with elevated fasting blood glucose, increased blood pressure, central obesity, decreased high-density lipoprotein, and increased triglyceride levels. Because of its growing incidence and prevalence and the effect that it has on developing other noncommunicable diseases, the importance gets even more value. The prediction and control of this disease in its early stages and in the most inexpensive way is of crucial need. Due to the role of chronic low-grade inflammation in metabolic syndrome, cytokines and inflammatory factors (like interleukin-6 and tumor necrosis factor-α) play a critical role on this phenomenon. The NLR is an inflammatory marker that has an unchallenging availability and has a reasonable cost. The NLR is related to obesity, type 2 diabetes, hypertension, and blood cholesterol levels. CONCLUSION: This suggests that there may be a relationship between the NLR and metabolic syndrome. The NLR, as a low-grade inflammation marker, indicates a positive relationship with central obesity. Also, studies indicate that the incidence of diabetes, its severity, and its control are related to the NLR. Hypertension and hyperlipidemia can both be noticed with higher NLRs. This rapid review assesses the association between metabolic syndrome and the NLR.
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Diabetes Mellitus Tipo 2 , Síndrome Metabólica , Biomarcadores , Diabetes Mellitus Tipo 2/metabolismo , Humanos , Linfócitos/metabolismo , Linfócitos/patologia , Síndrome Metabólica/metabolismo , Neutrófilos/metabolismo , Neutrófilos/patologiaRESUMO
Cancer is a primary cause of mortality around the world and imposes a significant physiological, psychological, and financial burden on patients. Lipids regulate cell cycle progression and affect cell proliferation, migration, and apoptosis. Therefore, alterations in serum lipid levels might contribute to carcinogenesis. In this article, we review the relationships between triglyceride (TG), total cholesterol, high-density lipoprotein cholesterol (HDL-C), and low-density lipoprotein cholesterol (LDL-C) levels and different types of cancer. Then, we examine the association between cancer and familial hypercholesterolemia. Finally, we evaluate the impact of statins on different types of cancer. Increased total cholesterol has been reported to increase cellular proliferation and angiogenesis in tumors and inhibit apoptosis. Increased LDL-C has been reported to induce inflammation and increase susceptibility to oxidative damage. HDL-C has anti-oxidation, anti-inflammatory, and antiproliferative properties. Increased levels of serum TG can induce oxidative stress and a chronic inflammatory state and therefore contribute to the proliferation and progression of cancer cells. Statins decrease downstream products of cholesterol synthesis that are crucial in cell proliferation and growth. Thus, lipid components can have prognostic value in cancer and management of serum lipid levels through lifestyle changes and medical therapy can be beneficial in cancer prevention and treatment.
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Background: Leishmania major is the etiologic agent of zoonotic cutaneous leishmaniasis in Iran, and glucantime injection is currently used for its treatment. This study aimed to evaluate the anti-leishmanial effect of topical Plantago psyllium and white vinegar in L. major infected BALB/c mice. Methods: Thirty infected mice were divided into five groups as follows: Group 1: treated with the combination of ovata powder and white vinegar, Group 2: treated with glucantime, Group 3: treated with white vinegar, Group 4: treated with the combination of ovata powder and water, and Group 5: without any treatment. All the groups were treated for 18 days. Lesion size was measured, and final smears were prepared for microscopic examination. Results: The findings indicated that the difference in the mean areas of the ulcers in all the groups before and after treatment was not significant, except for the second (glucantime) and third (vinegar) groups. Also, the results showed that in the first, second, third, and fourth group, 6 (60%), 4 (80%), 3 (60%), and 2 (40%) mice were healed, respectively. However, ulcers remained in all the five mice of the control group. Conclusion: The combination of ovata powder and white vinegar has been traditionally used to treat leishmanial lesions in Iran. It seems the most anti-leishmanial effect is related to vinegar and supported by Plantago. The route of treatment with this combination is very simple and painless in comparison with injection. Thus, further studies on this issue could help to design more effective and easy-to-use drugs.
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The COVID-19 pandemic has led to over 100 million infections and over 3 million deaths worldwide. Understanding its pathogenesis is crucial to guide prognostic and therapeutic implications. Viral infections are known to alter the lipid profile and metabolism of their host cells, similar to the case with MERS and SARS-CoV-2002. Since lipids play various metabolic roles, studying lipid profile alterations in COVID-19 is an inevitable step as an attempt to achieve better therapeutic strategies, as well as a potential prognostic factor in the course of this disease. Several studies have reported changes in lipid profile associated with COVID-19. The most frequently reported changes are a decline in serum cholesterol and ApoA1 levels and elevated triglycerides. The hyper-inflammatory state mediated by the Cytokine storm disturbs several fundamental lipid biosynthesis pathways. Virus replication is a process that drastically changes the host cell's lipid metabolism program and overuses cell lipid resources. Lower HDL-C and ApoA1 levels are associated with higher severity and mortality rates and with higher levels of inflammatory markers. Studies suggest that arachidonic acid omega-3 derivatives might help modulate hyper-inflammation and cytokine storm resulting from pulmonary involvement. Also, statins have been shown to be beneficial when administered after COVID-19 diagnosis via unclear mechanisms probably associated with anti-inflammatory effects and HDL-C rising effects.
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COVID-19 , Teste para COVID-19 , Humanos , Inflamação , Pandemias , SARS-CoV-2RESUMO
OBJECTIVE: The maternal immune response to paternal antigens is induced at insemination. We believe that pregnancy protective alloantibodies, such as anti-paternal cytotoxic antibody (APCA), may be produced against the paternal antigens in the context of stimulated immunity at insemination and that they increase during pregnancy. APCA is necessary for pregnancy. It is directed towards paternal human leucocyte antigens (HLAs) and has cytotoxic activity against paternal leucocytes. The present study aims to determine whether APCA is produced by the maternal peripheral blood mononuclear cells (PBMCs) in contact with the husband's spermatozoa and to evaluate the relation of APCA production with HLA class I and II expressions by spermatozoa in fertile couples. MATERIALS AND METHODS: This cross-sectional study included 30 fertile couples with at least one child. The maternal PBMCs were co-cultured with the husband's spermatozoa and the supernatant was assessed for the presence of IgG by ELISA. Cytotoxic activity of the supernatant on the husband's PBMCs was assessed by the complement-dependent cytotoxicity (CDC) assay. RESULTS: IgG was produced in all co-cultures, and the mean level of supernatant IgG was 669 ng/ml. The cytotoxic activity of the supernatant was observed in all the supernatant obtained from the co-cultures. The mean percentage of APCA in supernatant was 73.93%. CONCLUSION: Based on the results of this study it can be concluded that APCA may be a natural anti-sperm antibody (ASA), which can be produced during exposure to spermatozoa and may have some influence before pregnancy. Further research is required to determine the role of APCA before pregnancy.
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Background: Several animal studies have shown the roles of cytokines in regulating liver regeneration following liver resection (LR), which is a type of surgery designed to remove cancerous tumors from the liver. This study investigated how the expressions and serum levels of some pro- and anti-inflammatory cytokines in patients with hepatocellular carcinoma (HCC) were changed during LR. Methods: Liver tissues from 15 patients with HCC were collected and the levels of interleukin-6 (IL-6), tumor necrosis factor-alpha (TNF-α), IL-1α, IL-1 ß, IL-10, and transforming growth factor-beta1 (TGF-ß1) were assessed using real-time PCR assay at different times before and after LR. The serum values of TNF-α and IL-6 were also measured by ELISA. Results: After 60 and 90 minutes of LR, IL-6 gene expression was significantly increased (P < 0.001 - 0.05). The same trend was also observed in TNF-α expression after 90 minutes of LR (P < 0.01). No significant changes were observed in the expressions of IL-1α, IL-1ß, IL-10, and TGF-ß1 before and after LR. In addition, LR had significant effects on TNF-α and IL-6 serum levels (P < 0.05 - 0.0001). Conclusion: Our data provided further evidence to reveal that IL-6 and TNF-α cytokines are critical to improve liver regeneration.
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Carcinoma Hepatocelular , Neoplasias Hepáticas , Animais , Carcinoma Hepatocelular/cirurgia , Citocinas , Humanos , Neoplasias Hepáticas/cirurgia , Fator de Necrose Tumoral alfaRESUMO
PURPOSE: Progesterone-induced blocking factor (PIBF) is a protein secreted by lymphocytes exposed to progesterone (P4). P4 and PIBF have immunomodulatory effects on peripheral CD4+ T cells during normal pregnancy. Membrane progesterone receptors (mPRs) may correlate with the immunomodulatory properties of P4 on T cells. Variation in expression of mPRs may influence P4 regulatory performance during pregnancy. On the other hand, PIBF increases in pregnant normal women compared to women who have experienced abortion. The present study aimed to determine whether PIBF, in addition to having a direct influence on the immune system, can affect P4 performance through its effect on mPR expression. Such novel research findings demonstrate the importance of PIBF in the maintenance of pregnancy. METHODS: Isolated peripheral blood mononuclear cells (PBMCs) from 30 healthy women were stimulated with the mitogen phytohemagglutinin (PHA). Cells were either exposed to various concentrations of PIBF or had no exposure at all in a culture medium at 37 °C for 3 days. The mean fluorescence intensity (MFI) of mPRα and mPRß was evaluated using polyclonal and monoclonal antibodies on CD4+ T cells. RESULTS: PIBF was able to significantly increase mPR expression on the surface of peripheral CD4+ T cells (p ≤ 0.05). CONCLUSION: This study characterized the effects of PIBF on mPR expression on peripheral CD4+ T cells of healthy fertile women. Thus, a decrease in PIBF concentration during abnormal pregnancy can modulate mPR expression and regulatory performance of P4 on T cells. Future research into this issue is likely to open up a new understanding of the etiology of abortion.
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Linfócitos T CD4-Positivos/metabolismo , Proteínas da Gravidez/metabolismo , Receptores de Progesterona/metabolismo , Fatores Supressores Imunológicos/metabolismo , Aborto Espontâneo , Feminino , Humanos , Leucócitos Mononucleares , Gravidez , Progesterona/farmacologiaRESUMO
Autoimmune thyroid disease is the most common endocrine disorder during pregnancy. Thyroid autoantibodies (TAs) have been suggested to serve a role in implantation failure and spontaneous abortion. Until now, there are no data on the potential interaction of TAs with human reproductive organs. Here, we set out for the first time to test this hypothesis by studying the expression of thyroid peroxidase (TPO) at gene and protein level in human reproductive organs. Endometrial samples were taken from normal women, and placenta tissues were collected after full-term caesarian section. Expression of TPO messenger RNA (mRNA) was investigated by qRT-PCR. In addition, polyclonal anti-TPO antibodies were produced and the expression of TPO protein in mentioned tissues was evaluated by immunohistochemistry and Western blot analysis. The reactivity of anti-TPO antibody in human embryos was evaluated by immunofluorescent staining. For the first time, our study showed that TPO is expressed at gene and protein levels in endometrium and placenta. TPO expression was mainly localized to glandular and luminal epithelial cells in the endometrium. In placenta, the syncytiotrophoblasts and invasive trophoblast cells were the main cell types that expressed TPO protein. Specific band of approximately 110 kDa was observed in all endometrial and placental tissues by Western blot analysis. However, no expression of TPO protein was observed in human embryo. TPO expression in endometrium and placenta may explain higher frequency of abortion and infertility in patients with thyroid autoimmunity.
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Anticorpos/imunologia , Autoanticorpos/imunologia , Autoantígenos/metabolismo , Embrião de Mamíferos/metabolismo , Endométrio/metabolismo , Iodeto Peroxidase/metabolismo , Proteínas de Ligação ao Ferro/metabolismo , Placenta/metabolismo , Animais , Autoantígenos/genética , Autoantígenos/imunologia , Embrião de Mamíferos/imunologia , Endométrio/imunologia , Feminino , Seguimentos , Humanos , Iodeto Peroxidase/genética , Iodeto Peroxidase/imunologia , Proteínas de Ligação ao Ferro/genética , Proteínas de Ligação ao Ferro/imunologia , Placenta/imunologia , Gravidez , CoelhosRESUMO
OBJECTIVES: To compare the therapeutic effects of high-flow-oxygen-Therapy (HFT) and noninvasive-ventilation (NIV) for stabilizing chronic obstructive pulmonary disease during exacerbation. METHODS: In this randomized clinical trial at Masih-Daneshvari hospital, between July 2019 and Oct 2019, 30 exacerbated-COPD-patient with PaCO2 64.58 ± 11.61 mm Hg, Respiratory Rate 24.43 ± 2.75, and PH 7.31 ± 0.02 were divided into two groups, N = 15. By a simple randomized allocation, patients receive either NIV or HFT for 1 hour, and following a washout period of 30 minutes, they switched to the other treatment option. Arterial Blood Gas Parameters, as well as Respiratory Rate (RR), Dyspnea Score, Heart Rate (HR), and Oxygen Saturation (SO2 ), were compared before and after the intervention and between groups. RESULTS: Baseline patient characteristics were similar in the two groups. Pre and post-analysis revealed that in both groups, all improved significantly. After the first period, there was no difference in all parameters between groups except for SO2 which was significantly higher in HFT (%92.1 ± 1) than that of NIV (%89 ± 1), P = .001. Likewise, following the washout period, patients in HFT and NIV had a dyspnea score of 1.93 ± 0.7 and 2.73 ± 0.9, respectively, P = .01. No carryover-effect and was observed but the period effect was significant for some outcomes. A significant improvement in SO2 and HR was observed by HFT according to treatment effect by combining two periods' results. During the study, no side effects were reported. CONCLUSION: In this short-term study HFT appears feasible for patients with COPD exacerbation to reduce dyspnea score and improve respiratory distress.
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Ventilação não Invasiva , Doença Pulmonar Obstrutiva Crônica , Insuficiência Respiratória , Humanos , Pulmão , Oxigenoterapia , Doença Pulmonar Obstrutiva Crônica/terapia , Insuficiência Respiratória/terapiaRESUMO
BACKGROUND: Regarding to the increase of cancer deaths in recent years and disability of common therapies to eradicate cancers, as well as expansion of Natural Killer (NK) cell therapy, it seems so vital to find new useful therapies against cancers. Breast cancer is the second main cause of cancer death among women. As it is impossible for a majority of patients to receive NK cell therapy, an attempt was made to establish a low-cost and efficient method for expanding and activating NK cells against breast cancer cell line (MCF7). METHODS: NK cells were isolated from Peripheral Blood Mononuclear Cells (PBMCs) applying either MACS based NK cell enrichment kit or antibodies and complement as cytotoxic method. Then, the NK cells were cultured in Stem Cell Growth Medium (SCGM) with feeder layer (irradiated PBMCs) along with PHA or OKT3. IL-2, IL-15 and IL-21 were used to expand NK cells and finally their cytotoxic activity was investigated by flow cytometry. RESULTS: Highly pure NK cells were obtained and no significant difference between the two isolation methods was found. Using IL-2 plus IL-15, the number of NK cells increased up to100 fold after 16 days. No significant effect was observed after IL-21 treatment. CONCLUSION: Our data indicated that cytotoxicity method can be considered a low-cost alternative for NK cell isolation kits. It seems that culturing NK cells for 14 days in either PHA or OKT3 supplemented SCGM medium would be more effective than culturing for 16 days in the presence of IL-21.
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BACKGROUND & OBJECTIVE: Unexplained recurrent spontaneous abortion (URSA) is defined as an unknown cause of occurrence of three or more clinically detectable pregnancy losses before 20 weeks of gestation, but it occurs presumably as a result of the immune system dysfunctions. We supposed that the disruption of semen or spermatozoa might be responsible for the dysfunction of the immune system in women with URSA. Semen and spermatozoa (as antigens) induce female reproductive tract (FRT) immunity. This stimulated immunity is necessary for pregnancy occurrence. The disruption of semen or spermatozoa can be a result of altering a variety of surface molecules on spermatozoa, especially polymorphic human leukocyte antigen (HLA) molecules or antigens. Despite the importance of HLA antigens in reproduction, to the best of our knowledge, no one has studied the relation of HLA expression between spermatozoa and URSA. Therefore, this paper aims to assess this relation. METHODS: Semen samples were collected from 15 URSA couples and 20 normal couples. After purification of normal spermatozoa, the HLA class I and II expressions were evaluated by flow cytometry methods. RESULTS: Results showed that the expression of both HLA class I and II by spermatozoa, in URSA couples, was significantly less than the control couples. CONCLUSION: The decreased expression of polymorphic HLA class â and â ¡ by spermatozoa can be related to URSA occurrence.
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BACKGROUND: Genetic defects of pediatric-onset inflammatory bowel disease (IBD) provide critical insights into molecular factors controlling intestinal homeostasis. NOX1 has been recently recognized as a major source of reactive oxygen species (ROS) in human colonic epithelial cells. Here we assessed the functional consequences of human NOX1 deficiency with respect to wound healing and epithelial migration by studying pediatric IBD patients presenting with a stop-gain mutation in NOX1. METHODS: Functional characterization of the NOX1 variant included ROS generation, wound healing, 2-dimensional collective chemotactic migration, single-cell planktonic migration in heterologous cell lines, and RNA scope and immunohistochemistry of paraffin-embedded patient tissue samples. RESULTS: Using exome sequencing, we identified a stop-gain mutation in NOX1 (c.160C>T, p.54R>*) in patients with pediatric-onset IBD. Our studies confirmed that loss-of-function of NOX1 causes abrogated ROS activity, but they also provided novel mechanistic insights into human NOX1 deficiency. Cells that were NOX1-mutant showed impaired wound healing and attenuated 2-dimensional collective chemotactic migration. High-resolution microscopy of the migrating cell edge revealed a reduced density of filopodial protrusions with altered focal adhesions in NOX1-deficient cells, accompanied by reduced phosphorylation of p190A. Assessment of single-cell planktonic migration toward an epidermal growth factor gradient showed that NOX1 deficiency is associated with altered migration dynamics with loss of directionality and altered cell-cell interactions. CONCLUSIONS: Our studies on pediatric-onset IBD patients with a rare sequence variant in NOX1 highlight that human NOX1 is involved in regulating wound healing by altering epithelial cytoskeletal dynamics at the leading edge and directing cell migration.