Search results for: vascular microenvironment

Authors: Ming-Tzu Tsai, Jui-Ting Hsu, Chia-Chieh Lin, Feng-Tsai Chiang, Cheng-Chin Huang

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Far infrared (FIR), an invisible and short electromagnetic waves ranges from 6-14 μm also defines as the “growth ray.” Although the mechanism of FIR is still unknown, most data have suggested that FIR could accelerate the skin microcirculation by elevating the blood flow and nitric-oxide (NO) synthesis. In this present work, the effect of FIR irradiation and endothelial cell growth supplement (ECGS) on human umbilical vascular endothelial cells (HUVECs) was evaluated. To understand whether the cell viability and NO production of HUVECs affected by NO, cells with/without ECGS were treated in the presence or absence of L-NAME, an eNOS inhibitor. For FIR exposure, FIR-emitted ceramic powders consisted of a variety of well-mixed metal oxides were developed. The results showed that L-NAME did had a strong effect on the inhibition of NO production, especially in the ECGS-treated group. However, the cell viability of each group was rarely affected in the presence of L-NAME. Cells with the incubation of ECGS showed much higher cell viability compared to the control. Moreover, NO production of HUVECs exposed to FIR irradiation was significantly inhibited in the presence of L-NAME. It suggested that NO could play a role modulating the downstream signals of HUVECs during FIR exposure.

Keywords: far-infrared irradiation (FIR), nitric oxide (NO), endothelial nitric oxide synthase (eNOS), endothelial cell growth supplement (ECGS)

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Authors: Latha V. Prabhu, B.V. Murlimanju, P.J. Jiji, Mangala M. Pai

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Background: The capitate is the largest among the carpal bones. There exists no literature about the vascular foramina of the capitate bone. The objective of the present study was to investigate the morphology and number of the nutrient foramina in the cadaveric dried capitate bones of the Indian population. Methods: The present study included 59 capitate bones (25 right sided and 34 left sided) which were obtained from the gross anatomy laboratory of our institution. The bones were macroscopically observed for the nutrient foramina and the data was collected with respect to their number. The tabulation of the data and analysis were done. Results: All of our specimens (100%) exhibited the nutrient foramina over the non-articular and articular surfaces. The foramina were observed at the medial, lateral, palmar and dorsal surfaces of the capitate bones. The foramina were ranged from 6 to 23 in each capitate bone. In the medial surface, the foramina ranged from 1 to 6, lateral surface from 0 to 7, the foramina ranged between 0 and 5 in the palmar surface. However most of the foramina were located at the dorsal surface which ranged from 3 to 11. Conclusion: We believe that the present study has provided additional data about the nutrient foramina of the capitate bones. The data is enlightening to the orthopedic surgeon and would help in the hand surgeries. The knowledge about the foramina is also important to the radiologists to prevent the misinterpretation of the findings in the x ray and computed tomogram scan films. The foramina may mimick like erosions and ossicles. The morphological knowledge of the vasculature, their foramina of entry and number is required to understand the concepts in the avascular necrosis of the capitate.

Keywords: avascular necrosis, capitate, morphology, nutrient foramen

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Authors: Reetika Sharma, Lalit Tejwani, Himanshu Shekhar, Arun Singhvi

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Purpose: To describe a novel technique of trabeculectomy targeting triple modulation on wound healing to increase the overall success rate. Method: Ten eyes of 10 patients underwent trabeculectomy with subconjunctival mitomycin C (0.4mg/ml for 4 minutes) application combined with Ologen implantation subconjunctivally and subsclerally. Five of these patients underwent additional phacoemulsification with intraocular lens implantation. The Ologen implant was wet with 0.1 ml Bevacizumab. Result: All the eyes achieved target intraocular pressure (IOP), which was maintained until one year of follow-up. Two patients needed anterior chamber reformation at day two post surgery. One patient needed cataract surgery after four months of surgery and achieved target intraocular pressure on two topical antiglaucoma medicines. Conclusion: Vascular endothelial growth factor (VEGF) concentration has been seen to increase in the aqueous humor after filtration surgery. Ologen implantation helps in collagen remodelling, antifibroblastic response, and acts as a spacer. Bevacizumab augmented Ologen, in addition, targets the increased VEGF and helps in decreasing scarring. Anti-VEGF augmented Ologen in trabeculectomy with mitomycin C (MMC) hence appears to have encouraging short-term intraocular pressure control.

Keywords: ologen, anti-VEGF, trabeculectomy, scarring

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Authors: Juan Jose Filgueira, Daniela Londono-Serna, Liliana Maria Hoyos

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Carnation (Dianthus caryophyllus) is one of the most important products of exportation in the floriculture industry worldwide. Fusariosis is the disease that causes the highest losses on farms, in particular the one produced by Fusarium oxysporum f.sp. dianthi, called vascular wilt. Gene identification and metabolic routes of the genes that participate in the building of the plant response to Fusarium are some of the current targets in the carnation breeding industry. The techniques for the identifying of resistant genes in the plants, is the analysis of the transcriptome obtained during the host-pathogen interaction. In this work, we report the cell transcriptome of different varieties of carnation that present differential response from Fusarium oxysporum f.sp. dianthi attack. The cells of the different hybrids produced in the outbreeding program were cultured in vitro and elicited with the parasite in a dual culture. The isolation and purification of mRNA was achieved by using affinity chromatography Oligo dT columns and the transcriptomes were obtained by using Illumina NGS techniques. A total of 85,669 unigenes were detected in all the transcriptomes analyzed and 31,000 annotations were found in databases, which correspond to 36.2%. The library construction of genic expression techniques used, allowed to recognize the variation in the expression of genes such as Germin-like protein, Glycosyl hydrolase family and Cinnamate 4-hydroxylase. These have been reported in this study for the first time as part of the response mechanism to the presence of Fusarium oxysporum.

Keywords: Carnation, Fusarium, vascular wilt, transcriptome

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Authors: Saifullah K., Muhammad Naziruddin Saifullah, Muhammad N.

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The bio control potential and mechanism of P. chlamydosporia against Meloidogyne incognita was evaluated in the present study. Under invitro conditions, P. chlamydosporia was tested for parasitism of eggs and females of M. incognita. The results indicated that this fungus parasitized 87% eggs and 82% females. Culture filtrate (CF) of P. chlamydosporia was tested for its larvicide activity against M. incognita 2nd stage juvenile. The maximum mortality was 97.3% at 100% concentration of the culture filtrate while minimum mortality was 7.3% in 25% concentration after 24 hrs. The result of the pot experiment proved that P. chlamydosporia has reduced the incidence of RKN and improved all tested agronomic growth parameters. The treatment with inoculated M. incognita alone reduced plant height, fresh shoot, and fresh root weight by 44.7%, 29.8%, and 32.8% respectively over uninoculated healthy control. Histopathological studies on the interaction of Pochonia chlamydosporia and Meloidogyne incognita on tomato roots revealed anatomical changes among treatments. Less number of galls with small in size and scarcer abnormalities in the vascular cylinder was observed in plants inoculated with P. chlamydosporia and M. incognita than the plants treated with nematode only. The fungus was seen in in the intercellular spaces of cortical and epidermal cells while the vascular bundles of the plant remain intact, inoculated only with P. chlamydosporia. In the infected roots, many mature females were seen which feed on giant cells. The findings also revealed that control healthy plants were not affected and no histological changes were noted.

Keywords: histopathology, Pochonia chlamydosporia, Meloidogyne incognita, tomato

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Authors: Maria Stepankova, Lucie Vistejnova, Pavel Klein, Tereza Blassova, Marketa Slajerova, Radek Sedlacek, Martin Bartos, Jaroslav Chlupac

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Background: Clinical outcome benefits of the segment rib fracture surgical therapy are well known and follow from better stabilization of the chest wall. Despite this, some authors still incline to conservative therapy and point out to possible rib fracture healing failure in connection with the bone vascular supply disturbance caused by metal plate implantation. This suggestion met neither experimental nor clinical verification and remains the object of discussion. In our pilot study we investigated the titanium plate fixation effect on the rib fracture healing in porcine model and its histological, biomechanical and radiological aspects. Materials and Method: Two porcine models (experimental group) underwent the operative chest wall stabilization with a titanium plate implantation after osteotomy. Two other porcine models (control group) were treated conservatively after osteotomy. Three weeks after surgery, all animals were sacrificed, treated ribs were explanted and the histological analysis, µCT imaging and biomechanical testing of the calluses tissue were performed. Results: In µCT imaging, experimental group showed a higher cortical bone volume compared to the control group. Histological analysis using the non-decalcified bone tissue blocks demonstrated more maturated callus with higher newly-formed osseous tissue ratio in experimental group in comparison to controls. In contrast, no significant differences in bone blood vessels supply in both groups were observed. This finding suggests that the bone blood supply in experimental group was not impaired. Biomechanical analysis using 3-point bending test demonstrated significantly higher bending stiffness and the maximum force in experimental group. Conclusion: Based on our observation, it could be concluded, that the titanium plate fixation of the rib fractures leads to faster bone callus maturation whereas does not cause the vascular supply impairment after 3 weeks and thus has a beneficial effect on the rib fracture healing.

Keywords: bone vascular supply, chest wall stabilization, fracture healing, histological analysis, titanium plate implantation

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Authors: Marilou P. Angeles

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Seniors are a growing number of population all over the world, and they are getting sick with illnesses like diabetes, high blood, and high cholesterol. It is necessary to see the relationship of their physical illness and its effect on their quality of life. Having chronic illnesses also can affect the mood of the elderly; becoming cranky, lonely, not eating, etc. Therefore, there is a need to study the relationship of the quality of life of the elderly and the level of depression. Depression for elderly is known as late onset depression or vascular depression since it is tied to the vascular illnesses they are experiencing, although this is not homogeneous. There is heterogeneity in seniors. The purpose of the study is to determine how keep the satisfaction in life i.e., quality of life of seniors, as long as possible. This study was made in 4 barangays in Longos, Potrero, Tonsuya and Catmon, in Malabon, Metro Manila, Philippines. These Filipino seniors are availing of free medicines for their diabetes, high blood, and high cholesterol ailments in the barangay health centers, given freely by the Department of Health. Two instruments were used; quality of life (CASP-19) and patient health questionnaire(PHQ-19). The quality of life questionnaire was based on the theory of Abraham Maslow, human: beings are motivated to action by needs, starting from the lowest, physiological to the highest self-transcendence. Severity of depression is determined by PHQ-9, and according to the unified model of depression by Aaron Beck and Kurt B. Bredemeier, depression happens when a person cannot cope with life has not able to satisfy his needs as a person. The Pearson R correlation was used to determine the significance of the relationship between quality of life and depression. Finding is there is negative relationship between quality of life and depression. It means that a high value of quality of life lowers or minimizes depression. CASP-19 found that the Filipino elderly were in control, independent, enjoying their lives even if they are poor, and this is shown by the significant results. Self-transcendence, a need to give back to others, is important for Filipino elderly. Although the seniors have difficulty with money and they were affected by their illnesses, they are full of optimism, they are ignoring their physical pain because they are focusing on helping their loved ones (i.e., self-transcendence), their children and grandchildrenothers, and if problems come, they are resilient accepting of the challenges, because they have strong faith in God. They are also having pleasures interacting with their friends and neighbors who, like them, have the same health problems. And these two coping strategies for the elderlies allow them to live a meaningful life, a life high in quality. Thus, where there is high quality of life, there is none or minimal depression. Recommendation for future study is finding the relationship of spirituality to quality of life of seniors.

Keywords: CASP-19, depression, quality of life, PHQ-9, senior citizen

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Authors: Abhishek Sharma, Amit Gupta, Manish Singhal

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Background: The management of patients with penetrating neck injuries in the prehospital setting and in the emergency department has evolved with regard to the use of multidetector computed tomographic (MDCT) imaging. Hence, there is a shift in the management of neck injuries from mandatory exploration in certain anatomic areas to more conservative approach using imaging and so-called “no zone approach”. Objective: To study the no zone approach in the management of penetrating neck injury using routine imaging in all stable patients. Methods: 137 patients with penetrating neck injury attending emergency department of level 1 trauma centre at AIIMS between 2008–2014 were retrospectively analysed. All hemodynamically stable patients were evaluated using CT scanning. Results: Stab injury is most common (55.91%) mode of pni in civilian population followed by gunshot(18.33%). The majority of patients could be managed with imaging and close observation. 39 patients (28.46%) required operative intervention. The most common indication for operative intervention was vascular followed by airway injury manifesting as hemodynamic destabilisation.There was no statistical difference between the zonal distribution of injuries in patients managed conservatively and those taken to OR. Conclusions: Study shows that patients with penetrating neck trauma who are haemodynamically stable and exhibit no “hard signs” of vascular injury or airway injury may be evaluated initially by MDCT imaging even when platysma violation is present. “No Zone” policy may be superior to traditional zone wise management.

Keywords: penetrating neck injury, zone approach, CT scanning, multidetector computed tomographic (MDCT)

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Authors: Diana Islam, Juan Fang, Vito Fanelli, Bing Han, Julie Khang, Jianfeng Wu, Arthur S. Slutsky, Haibo Zhang

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Introduction: MSC delivery in preclinical models of ARDS has demonstrated significant improvements in lung function and recovery from acute injury. However, the role of MSC delivery in ARDS associated pulmonary fibrosis is not well understood. Some animal studies using bleomycin, asbestos, and silica-induced pulmonary fibrosis show that MSC delivery can suppress fibrosis. While other animal studies using radiation induced pulmonary fibrosis, liver, and kidney fibrosis models show that MSC delivery can contribute to fibrosis. Hypothesis: The beneficial and deleterious effects of MSC in ARDS are modulated by the lung microenvironment at the time of MSC delivery. Methods: To induce ARDS a two-hit mouse model of Hydrochloric acid (HCl) aspiration (day 0) and mechanical ventilation (MV) (day 2) was used. HCl and injurious MV generated fibrosis within 14-28 days. 0.5x106 mouse MSCs were delivered (via both intratracheal and intravenous routes) either in the active inflammatory phase (day 2) or during the remodeling phase (day 14) of ARDS (mouse fibroblasts or PBS used as a control). Lung injury accessed using inflammation score and elastance measurement. Pulmonary fibrosis was accessed using histological score, tissue collagen level, and collagen expression. In addition alveolar epithelial (E) and mesenchymal (M) marker expression profile was also measured. All measurements were taken at day 2, 14, and 28. Results: MSC delivery 2 days after HCl exacerbated lung injury and fibrosis compared to HCl alone, while the day 14 delivery showed protective effects. However in the absence of HCl, MSC significantly reduced the injurious MV-induced fibrosis. HCl injury suppressed E markers and up-regulated M markers. MSC delivery 2 days after HCl further amplified M marker expression, indicating their role in myofibroblast proliferation/activation. While with 14-day delivery E marker up-regulation was observed indicating their role in epithelial restoration. Conclusions: Early MSC delivery can be protective of injurious MV. Late MSC delivery during repair phase may also aid in recovery. However, early MSC delivery during the exudative inflammatory phase of HCl-induced ARDS can result in pro-fibrotic profiles. It is critical to understand the interaction between MSC and the lung microenvironment before MSC-based therapies are utilized for ARDS.

Keywords: acute respiratory distress syndrome (ARDS), mesenchymal stem cells (MSC), hydrochloric acid (HCl), mechanical ventilation (MV)

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Authors: Hansraj Riteesh Bookun, Emily Maree Stevens, Jarryd Leigh Solomon, Anthony Chan

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Objective: This was a retrospective cross-sectional study evaluating the safety and efficacy of femoral endarterectomy using the eversion technique with transection as opposed to the conventional endarterectomy technique with either vein or synthetic patch arterioplasty. Methods: Between 2010 to mid 2017, 19 patients with mean age of 75.4 years, underwent eversion femoral endarterectomy with transection by a single surgeon. There were 13 males (68.4%), and the comorbid burden was as follows: ischaemic heart disease (53.3%), diabetes (43.8%), stage 4 kidney impairment (13.3%) and current or ex-smoking (73.3%). The indications were claudication (45.5%), rest pain (18.2%) and tissue loss (36.3%). Results: The technical success rate was 100%. One patient required a blood transfusion following bleeding from intraoperative losses. Two patients required blood transfusions from low post operative haemogloblin concentrations – one of them in the context of myelodysplastic syndrome. There were no unexpected returns to theatre. The mean length of stay was 11.5 days with two patients having inpatient stays of 36 and 50 days respectively due to the need for rehabilitation. There was one death unrelated to the operation. Conclusion: The eversion technique with transection is safe and effective with low complication rates and a normally expected length of stay. It poses the advantage of not requiring a synthetic patch. This technique features minimal extraneous dissection as there is no need to harvest vein for a patch. Additionally, future endovascular interventions can be performed by puncturing the native vessel. There is no change to the femoral bifurcation anatomy after this technique. We posit that this is a useful adjunct to the surgeon’s panoply of vascular surgical techniques.

Keywords: endarterectomy, eversion, femoral, vascular

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Authors: Irina E. Sukovataya, Oleg S. Sutormin, Valentina A. Kratasyuk

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Effect of viscosity of media on kinetic parameters of the coupled enzyme system NADH:FMN-oxidoreductase–luciferase was investigated with addition of organic solvents (glycerol and sucrose), because bioluminescent enzyme systems based on bacterial luciferases offer a unique and general tool for analysis of the many analytes and enzymes in the environment, research, and clinical laboratories and other fields. The possibility of stabilization and increase of activity of the coupled enzyme system NADH:FMN-oxidoreductase–luciferase activity in vicious aqueous-organic mixtures have been shown.

Keywords: coupled enzyme system of bioluminescence bacteria NAD(P)H:FMN-oxidoreductase–luciferase, glycerol, stabilization of enzymes, sucrose

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Authors: Haaris Khan, Farhad Udwadia

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South Asians are the fastest-growing immigrant population in Canada and are 3-4 times more likely to develop diabetes. In a primary care setting, language barriers continue to persist as a prominent obstacle when delivering crucial health information. Given the abundance of languages in the South Asian community and the varying levels of English fluency, there is compelling evidence that these language barriers can adversely impact health outcomes. The microvascular and macrovascular complications of poor diabetic management are well established and universally recognized. However, these are often difficult concepts to grasp for even individuals fluent in English. In order to lessen the burden of language barriers, we developed a comprehensive guide in various languages that discuss the complications and screening guidelines for diabetic and prediabetic patients. The guide is presented in the form of a pamphlet, with an electronic version being constructed as well, that provides basic information on diabetic retinopathy, neuropathy and nephropathy as well as the screening recommendations. We also conducted a review of the literature around the topic and incorporated our findings into our project. Our goal is for primary care physicians to have this resource and to be able to provide the link or pamphlet to patients in need. Our presentation also provides a comprehensive overview of some of the other barriers that individuals in the South Asian community face when seeking care. Given the staggering number of individuals in the South Asian community with diabetes and the morbidity and mortality associated with diabetes and its complications, effective community-specific strategies are needed to mitigate the potential consequences of poor diabetes management.

Keywords: diabetes, patient education, ophthalmology, vascular surgery

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Authors: S. Jaramillo, Y. Montoya, W. Agudelo, J. Bustamante

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Cardiovascular diseases (CVD) are related to affectations of the heart and blood vessels, within these are pathologies such as coronary or peripheral heart disease, caused by the narrowing of the vessel wall (atherosclerosis), which is related to the accumulation of Low-Density Lipoproteins (LDL) in the arterial walls that leads to a progressive reduction of the lumen of the vessel and alterations in blood perfusion. Currently, the main therapeutic strategy for this type of alteration is drug treatment with statins, which inhibit the enzyme 3-hydroxy-3-methyl-glutaryl-CoA reductase (HMG-CoA reductase), responsible for modulating the rate of cholesterol production and other isoprenoids in the mevalonate pathway. This enzyme induces the expression of LDL receptors in the liver, increasing their number on the surface of liver cells, reducing the plasma concentration of cholesterol. On the other hand, when the blood vessel presents stenosis, a surgical procedure with vascular implants is indicated, which are used to restore circulation in the arterial or venous bed. Among the materials used for the development of vascular implants are Dacron® and Teflon®, which perform the function of re-waterproofing the circulatory circuit, but due to their low biocompatibility, they do not have the ability to promote remodeling and tissue regeneration processes. Based on this, the present research proposes the development of a hydrolyzed collagen and polyethylene oxide electrospun membrane reinforced with medium and high-intensity statins, so that in future research it can favor tissue remodeling processes from its microarchitecture.

Keywords: atherosclerosis, medium and high-intensity statins, microarchitecture, electrospun membrane

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Authors: Faycal Boughalleb, Raoudha Abdellaoui

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The effect of NaCl stress on root and leaf anatomy was investigated in three Astragalus species grown in 0-300 mM NaCl for 30 days under greenhouse conditions. Root cross section and cortex thickness was reduced under salt stress in both species while A. tenuifolius showed thinner cortex and the root cross section was unchanged. The epidermis stele thickness was unaffected by salinity in A. armatus and A. tenuifolius and was reduced in A. mareoticus with smaller xylem vessel size. In addition, vessel density and wall thickness of xylem was increased under salt conditions in the studies species. The entire lamina and mesophyll of the three species were thinner in salt-stressed plants. A. armatus and A. tenuifolius showed the higher thickness with increased size of the lower epidermis. NaCl (300 mM) reduced leaf water content by 41.5 % in A. mareoticus while it was unchanged in the other species. The size of the vascular bundle increased under salinity in A. tenuifolius leaves and it was unchanged in the other ones. A longer distance between leaf vascular bundle was occurred in A. mareoticus. The effects of NaCl on root and leaf ultrastructure are discussed in relation to the degree of salt resistance of these species. The unchanged biomass production under salt stress confirmed the higher tolerance oft A. tenuifolius to salinity. A. armatus was moderately salt tolerant with decrease of biomass production by 14.2 % while A. mareoticus was considered as salt sensitive plant when the decrease in biomass production reached 56.8%.

Keywords: Astragalus species, leaf ultrastructure, root anatomy, salt stress

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Authors: Ainur Sharip, Jose E. Perez, Nouf Alsharif, Aldo I. M. Bandeas, Enzo D. Fabrizio, Timothy Ravasi, Jasmeen S. Merzaban, Jürgen Kosel

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Bone marrow-derived human mesenchymal stem cells (MSCs) are attractive candidates for tissue engineering and regenerative medicine, due to their ability to differentiate into osteoblasts, chondrocytes or adipocytes. Differentiation is influenced by biochemical and biophysical stimuli provided by the microenvironment of the cell. Thus, altering the mechanical characteristics of a cell culture scaffold can directly influence a cell’s microenvironment and lead to stem cell differentiation. Mesenchymal stem cells were cultured on densely packed, vertically aligned magnetic iron nanowires (NWs) and the effect of NWs on the cell cytoskeleton rearrangement and differentiation were studied. An electrochemical deposition method was employed to fabricate NWs into nanoporous alumina templates, followed by a partial release to reveal the NW array. This created a cell growth substrate with free-standing NWs. The Fe NWs possessed a length of 2-3 µm, with each NW having a diameter of 33 nm on average. Mechanical stimuli generated by the physical movement of these iron NWs, in response to a magnetic field, can stimulate osteogenic differentiation. Induction of osteogenesis was estimated using an osteogenic marker, osteopontin, and a reduction of stem cell markers, CD73 and CD105. MSCs were grown on the NWs, and fluorescent microscopy was employed to monitor the expression of markers. A magnetic field with an intensity of 250 mT and a frequency of 0.1 Hz was applied for 12 hours/day over a period of one week and two weeks. The magnetically activated substrate enhanced the osteogenic differentiation of the MSCs compared to the culture conditions without magnetic field. Quantification of the osteopontin signal revealed approximately a seven-fold increase in the expression of this protein after two weeks of culture. Immunostaining staining against CD73 and CD105 revealed the expression of antibodies at the earlier time point (two days) and a considerable reduction after one-week exposure to a magnetic field. Overall, these results demonstrate the application of a magnetic NW substrate in stimulating the osteogenic differentiation of MSCs. This method significantly decreases the time needed to induce osteogenic differentiation compared to commercial biochemical methods, such as osteogenic differentiation kits, that usually require more than two weeks. Contact-free stimulation of MSC differentiation using a magnetic field has potential uses in tissue engineering, regenerative medicine, and bone formation therapies.

Keywords: cell substrate, magnetic nanowire, mesenchymal stem cell, stem cell differentiation

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Authors: Kajori Banerjee, Laxmi Kant Dwivedi

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India is a part of the ongoing globalization, contemporary convergence, industrialization and technical advancement that is taking place world-wide. Some of the manifestations of this evolution is rapid demographic, socio-economic, epidemiological and health transition. There has been a considerable increase in non-communicable diseases due to change in lifestyle. This study aims to assess the direction of burden of disease and compare the pressure of infectious diseases against cardio-vascular, endocrine, metabolic and nutritional diseases. The change in prevalence in a ten-year period (2004-2014) is further decomposed to determine the net contribution of various socio-economic and demographic covariates. The present study uses the recent 71st (2014) and 60th (2004) rounds of National Sample Survey. The pressure of infectious diseases against cardio-vascular (CVD), endocrine, metabolic and nutritional (EMN) diseases during 2004-2014 is calculated by Prevalence Rates (PR), Hospitalization Rates (HR) and Case Fatality Rates (CFR). The prevalence of non-communicable diseases are further used as a dependent variable in a logit regression to find the effect of various social, economic and demographic factors on the chances of suffering from the particular disease. Multivariate decomposition technique further assists in determining the net contribution of socio-economic and demographic covariates. This paper upholds evidences of stagnation of the burden of communicable diseases (CD) and rapid increase in the burden of non-communicable diseases (NCD) uniformly for all population sub-groups in India. CFR for CVD has increased drastically in 2004-2014. Logit regression indicates the chances of suffering from CVD and EMN is significantly higher among the urban residents, older ages, females, widowed/ divorced and separated individuals. Decomposition displays ample proof that improvement in quality of life markers like education, urbanization, longevity of life has positively contributed in increasing the NCD prevalence rate. In India’s current epidemiological phase, compression theory of morbidity is in action as a significant rise in the probability of contracting the NCDs over the time period among older ages is observed. Age is found to play a vital contributor in increasing the probability of having CVD and EMN over the study decade 2004-2014 in the nationally representative sample of National Sample Survey.

Keywords: cardio-vascular disease, case-fatality rate, communicable diseases, hospitalization rate, multivariate decomposition, non-communicable diseases, prevalence rate

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Authors: Mihai-Ionut Firescu, Mark A. P. Carson

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We present a case of inferior vena cava agenesis (IVCA) associated with bilateral deep venous thrombosis (DVT) in a patient with Beckwith-Wiedemann syndrome (BWS). In adult patients with BWS presenting with bilateral lower limb oedema, specific aetiological factors should be considered. These include cardiomyopathy and intraabdominal tumours. Congenital malformations of the IVC, through causing relative venous stasis, can lead to lower limb oedema either directly or indirectly by favouring lower limb venous thromboembolism; however, they are yet to be reported as an associated feature of BWS. Given its life-threatening potential, the prompt initiation of treatment for bilateral DVT is paramount. In BWS patients, however, this can prove more complicated. Due to overgrowth, the above-average birth weight can continue throughout childhood. In this case, the patient’s weight reached 170 kg, impacting on anticoagulation choice, as direct oral anticoagulants have a limited evidence base in patients with a body mass above 120 kg. Furthermore, the presence of IVCA leads to a long-term increased venous thrombosis risk. Therefore, patients with IVCA and bilateral DVT warrant specialist consideration and may benefit from multidisciplinary team management, with hematology and vascular surgery input. Conclusion: Here, we showcased a rare cause for bilateral lower limb oedema, respectively bilateral deep venous thrombosis complicating IVCA in a patient with Beckwith-Wiedemann syndrome. The importance of this case lies in its novelty, as the association between IVC agenesis and BWS has not yet been described. Furthermore, the treatment of DVT in such situations requires special consideration, taking into account the patient’s weight and the presence of a significant, predisposing vascular abnormality.

Keywords: Beckwith-Wiedemann syndrome, bilateral deep venous thrombosis, inferior vena cava agenesis, venous thromboembolism

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Authors: Liang Ning, Chung Hau Yin

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Angiogenesis is the process of new blood vessel development. It has been recognized as a therapeutic target for blocking cancer growth four decades ago. Vascular sprouting is initiated by pro-angiogenic factors. Vascular endothelial cell growth factor (VEGF) plays a central role in angiogenic initiation, many patients with cancer or ocular neovascularization have been benefited from anti-VEGF therapy. Emerging approaches impacting in the later stages of vessel remodeling and maturation are expected to improve clinical efficacy. TIE receptor as well as the corresponding angiopoietin ligands, were identified as another endothelial cell specific receptor tyrosine kinase signaling system. Much efforts were made to reduce the activity of angiopoietin-TIE receptor axis. Two eudesmane-type sesquiterpenes from laggera alata, namely, 15-dihydrocostic acid and ilicic acid were found with strong anti-angiogenic properties in zebrafish model. Meanwhile, the mRNA expression levels of VEGFR2 and TIE2 pathway related genes were down-regulated in the sesquiterpenes treated zebrafish embryos. Besides, in human umbilical vein endothelial cells (HUVECs), the sesquiterpenes have the ability to inhibit VEGF-induced HUVECs proliferation and migration at non-toxic concentration. Moreover, angiopoietin-2 induced TIE2 phosphorylation was inhibited by the sesquiterpenes, the inhibitory effect was detected in angiopoietin-1 induced HUVECs proliferation as well. Thus, we hypothesized the anti-angiogenic activity of the compounds may via the inhibition of VEGF and TIE2 related pathways. How the compounds come into play as the pathways inhibitors need to be evaluated in the future.

Keywords: Laggera alata, eudesmane-type sesquiterpene, anti-angiogenesis, VEGF, angiopoietin, TIE2

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Authors: Pagadaplly Girish, P. V. Manohar

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Introduction: Management of distal femur fractures is challenging. Recently, treatment has evolved towards indirect reduction and minimally invasive techniques. Objectives: To assess the fracture union and functional outcome following open reduction and internal fixation of distal femur fractures with locking compression plate and to achieve restoration of the anatomical alignment of fracture fragments and stable internal fixation. Methodology: Patients with distal femur fracture treated by locking compression during Oct 2011 to April 2013 were assessed prospectively. Patients below 18 years and those with neuro-vascular deficits were excluded. Age, sex of the patient, type of fracture, mechanism of injury, type of implant used, operative time and postoperative complications were analysed. The Neer’s scale was used to assess the outcome of the patients. Results: The total number of patients was 30; 28 males and 2 females; mean age was 41.53 years. Road traffic accidents were the major causes of injury followed by falls. The average duration of hospital stay was 21.3 days. The overall complication rate note was 23.33%. The mean range of movement around the knee joint after 6 months of follow-up was 114.330. The average time for the radiological union was 14 weeks. Excellent to good results were noted in 26 patients (86.6%) and average to poor results were observed in 4 (13.33%) patients. Conclusions: The locking compression plate gives a rigid fixation for the fracture. It also provides a good purchase in osteoporotic bones. LCP is simple and a reliable implant appropriate for fixation of femoral fractures with promising results.

Keywords: distal femur fractures, locking compression plate, Neer’s criteria, neuro-vascular deficits

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Authors: Dedy Pratama, Akhmadu Muradi, Hilman Ibrahim, Raden Suhartono, Alexander Jayadi Utama, Patrianef Darwis, S. Dwi Anita, Luluk Yunaini, Kemas Dahlan

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Introduction: Vascular endothelial growth factor (VEGF) gene shows association with various angiogenesis conditions including Diabetic Foot Ulcer (DFU) disease. In this study, we performed this study to examine VEGF gene polymorphism associated with DFU. Methods: Case-control study of polymorphism of VEGF gene +405 C>G and -460 T>C, of diabetes mellitus (DM) type 2 with Diabetic Foot Ulcer (DFU) in Cipto Mangunkusumo National Hospital (RSCM) Jakarta from June to December 2016. Results: There were 203 patients, 102 patients with DFU and 101 patients without DFU. Forty-nine point 8 percent of total samples is male and 50,2% female with mean age 56,06 years. Distribution of the wild-type genotype VEGF +405 C>G wild type CC was found in 6,9% of respondents, the number of mutant heterozygote CG was 69,5% and mutant homozygote GG was 19,7%. Cumulatively, there were 6,9% wild-type and 85,2% mutant and 3,9% of total blood samples could not be detected on PCR-RFLP. Distribution of VEGF allele +405 C>G C alleles were 43% and G alleles were 57%. Distribution of genotype from VEGF gene -460 T>C is wild type TT 42,9%, mutant heterozygote TC 37,9% and mutant homozygote CC 13,3%. Cumulatively, there were 42,9% wild-type and 51% mutant type. Distribution of VEGF -460 T>C were 62% T allele and 38% C allele. Conclusion: In this study we found the distribution of alleles from VEGF +405 C>G is C 43% and G 57% and from VEGF -460 T>C; T 62% and C 38%. We propose that G allele in VEGF +405 C>G can act as a protective allele and on the other hands T allele in VEGF -460 T>C could be acted as a risk factor for DFU in diabetic patients.

Keywords: diabetic foot ulcer, diabetes mellitus, polymorphism, VEGF

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Authors: Biju Lukose

Abstract:

The objective of the study was to identify the isolated and combined effect of multi-media computer assisted coaching and traditional coaching on selected motor ability component and physiological variables among sports school basketball players. Forty male basketball players aged between 14 to 18 years were selected randomly. They were divided into four groups of three experimental and one control. Isolated multi-media computer assisted coaching, isolated traditional coaching and combined coaching (multimedia computer assisted coaching and traditional coaching) are the three experimental groups. All the three experimental groups were given coaching for 24 weeks and control group were not allowed to participate in any coaching programme. The subjects were tested dependent variables such as speed and cardio vascular endurance; at the beginning (pre-test) in middle 12 week (mid-test) and after the coaching 24 week (post-test). The coaching schedule was for a period of 24 weeks. The data were collected two days before and after the coaching schedule and mid test after the 12 weeks of the coaching schedule. The data were analysed by applying ANCOVA and Scheffe’s Post hoc test. The result showed that there were significant changes in dependent variables such as speed and cardio vascular endurance. The results of the study showed that combined coaching (multimedia computer assisted coaching and traditional coaching) is more superior to traditional coaching and multimedia computer assisted coaching groups and no significant change in speed in the case of isolated multimedia computer assisted coaching group.

Keywords: computer, computer-assisted coaching, multimedia coaching, traditional coaching

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Authors: Rehab F. Abdel-Rahman, Sally A. El Awdan, Rehab R. Hegazy, Dina F. Mansour, Hanan A. Ogaly, Marwan Abdelbaset

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Disorders of the cerebral circulation are the leading cause of numerous neurological and psychiatric illnesses. The transient middle cerebral artery occlusion model (MCAO) is considered to be a reliable and reproducible rodent model of cerebral ischemia. The purpose of the current study was to examine the neuroprotective effects of Crocus sativus (saffron) in a rat model of left middle cerebral artery MCAO. Male Wistar rats were anesthetized and subjected to 1 h of MCAO followed by 48 h reperfusion or sham surgery. One group of the ischemia operated animals was kept as left brain ischemia/reperfusion (I/R). Another 2 operated groups received saffron extract (100 or 200 mg/kg, i.p) four times (60 min before the surgery, during the surgery, and on days 1 and 2 after the occlusion). During the experiment, behavioral tests were performed. After 72 h the animals were euthanized and their left brain hemispheres were used in the biochemical, histopathological, and immunohistochemical studies. Saffron administration revealed an improvement in I/R-induced alteration of locomotor balance and coordination ability of rats. Moreover, saffron decreased the brain content of malondialdehyde, nitric oxide, brain natriuretic peptide and vascular endothelial growth factor with significant increase of reduced glutathione. Immunohistochemical evaluation of caspase-3 and Bax protein expression revealed reduction in I/R-enhanced apoptosis in saffron treated rats. In conclusion, saffron treatment decreases ischemic brain injury in association with inhibition of apoptotic and oxidative cell death in a dose dependent manner.

Keywords: caspase-3, cerebral ischemia, Crocus sativus, rats, vascular endothelial growth factor

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Authors: Hajji Lobna, Chattaoui Mayssa, Regaieg Hajer, M'Hamdi-Boughalleb Naima, Rhouma Ali, Horrigue-Raouani Najet

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In this study, three local isolates of Trichoderma (Tr1: T. viride, Tr2: T. harzianum and Tr3: T. asperellum) were isolated and evaluated for their biocontrol effectiveness under in vitro conditions and in greenhouse. In vitro bioassay revealed a biopotential control against Fusarium oxysporum f. sp. radicis lycopersici and Meloidogyne javanica (RKN) separately. All species of Trichoderma exhibited biocontrol performance and (Tr1) Trichoderma viride was the most efficient. In fact, growth rate inhibition of Fusarium oxysporum f. sp. radicis lycopersici (FORL) was reached 75.5% with Tr1. Parasitism rate of root-knot nematode was 60% for juveniles and 75% for eggs with the same one. Pots experiment results showed that Tr1 and Tr2, compared to chemical treatment, enhanced the plant growth and exhibited better antagonism against root-knot nematode and root-rot fungi separated or combined. All Trichoderma isolates revealed a bioprotection potential against Fusarium oxysporum f. sp. radicis lycopersici. When pathogen fungi inoculated alone, Fusarium wilt index and browning vascular rate were reduced significantly with Tr1 (0.91, 2.38%) and Tr2 (1.5, 5.5%), respectively. In the case of combined infection with Fusarium and nematode, the same isolate of Trichoderma Tr1 and Tr2 decreased Fusarium wilt index at 1.1 and 0.83 and reduced the browning vascular rate at 6.5% and 6%, respectively. Similarly, the isolate Tr1 and Tr2 caused maximum inhibition of nematode multiplication. Multiplication rate was declined at 4% with both isolates either tomato infected by nematode separately or concomitantly with Fusarium. The chemical treatment was moderate in activity against Meloidogyne javanica and Fusarium oxysporum f. sp. radicis lycopersici alone and combined.

Keywords: trichoderma spp., meloidogyne javanica, Fusarium oxysporum f.sp.radicis lycopersici, biocontrol

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Authors: Anita Conti, Riccardo Ossanna, Lindsey A. Quintero, Giamaica Conti, Andrea Sbarbati

Abstract:

Ischemia/reperfusion (IR) injury induces muscle fiber atrophy and skeletal muscle fiber death with subsequently functionality loss. The heterogeneous pool of cells, especially mesenchymal stem cells, contained in the stromal vascular fraction (SVF) of adipose tissue could promote muscle fiber regeneration. To prevent SVF dispersion, it has been proposed the use of injectable biopolymers that work as cells carrier. A significant element of the extracellular matrix is hyaluronic acid (HA), which has been widely used in regenerative medicine as a cell scaffold given its biocompatibility, degradability, and the possibility of chemical functionalization. Connective tissue micro-fragments enriched with SVF obtained from mechanical disaggregation of adipose tissue were evaluated for IR muscle injury regeneration using low molecular weight HA as a scaffold. IR induction. Hindlimb ischemia was induced in 9 athymic nude mice through the clamping of the right quadriceps using a plastic band. Reperfusion was induced by cutting the plastic band after 3 hours of ischemic period. Contralateral (left) muscular tissue was used as healthy control. Treatment. Twenty-four hours after the IR induction, animals (n=3) were intramuscularly injected with 100 µl of SVF mixed with HA (SVF-HA). Animals treated with 100 µl of HA (n=3) and 100 µl saline solution (n=3) were used as control. Treatment monitoring. All animals were in vivo monitored by magnetic resonance imaging (MRI) at 5, 7, 14 and 18 days post-injury (dpi). High-resolution morphological T2 weighed, quantitative T2 map and Dynamic Contrast-Enhanced (DCE) images were acquired in order to assess the regenerative potential of SVF-HA treatment. Ex vivo evaluation. After 18 days from IR induction, animals were sacrificed, and the muscles were harvested for histological examination. At 5 dpi T2 high-resolution MR images clearly reveal the presence of an extensive edematous area due to IR damage for all groups identifiable as an increase of signal intensity (SI) of muscular and surrounding tissue. At 7 dpi, animals of the SVF-HA group showed a reduction of SI, and the T2relaxation time of muscle tissue of the HA-SVF group was 29±0.5ms, comparable with the T2relaxation time of contralateral muscular tissue (30±0.7ms). These suggest a reduction of edematous overflow and swelling. The T2relaxation time at 7dpi of HA and saline groups were 84±2ms and 90±5ms, respectively, which remained elevated during the rest of the study. The evaluation of vascular regeneration showed similar results. Indeed, DCE-MRI analysis revealed a complete recovery of muscular tissue perfusion after 14 dpi for the SVF-HA group, while for the saline and HA group, controls remained in a damaged state. Finally, the histological examination of SVF-HA treated animals exhibited well-defined and organized fibers morphology with a lateralized nucleus, similar to contralateral healthy muscular tissue. On the contrary, HA and saline-treated animals presented inflammatory infiltrates, with HA slightly improving the diameter of the fibers and less degenerated tissue. Our findings show that connective tissue micro-fragments enriched with SVF induce higher muscle homeostasis and perfusion restoration in contrast to control groups.

Keywords: ischemia/reperfusion injury, regenerative medicine, resonance imaging, stromal vascular fraction

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Authors: Dina Atmani-Kilani, Farah Yous, Djebbar Atmani

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The etiology of peptic ulcer is closely related to stress, excessive consumption of nonsteroidal anti-inflammatory drugs, or ethanol. Clematis flammula (Ranunculaceae) is a medicinal plant widely used by rural populations to treat inflammatory disorders. This study was designed to assess the gastroprotective potential of C. flammula extracts. Gastric ulcer was induced by stress, indomethacin, HCl / ethanol, and absolute ethanol on NMRI-type mice. The antioxidant potency of the ethanolic extract of Clematis flammula (EECF) was evaluated on catalase (CAT), superoxide dismutase (SOD), glutathione peroxidase (GPx) activities. Glutathione (GSH) and malonaldehyde (MDA) levels were also quantified. The anti-inflammatory potential was evaluated through the effect of EECF on myeloperoxidase activity (MPO) and vascular permeability. Complementary tests concerning the quantification of mucus levels, gastric motility, inhibition of ATPase H+/K+activity, as well as a histopathological study were also undertaken to explore the mechanism of action of the EECF. The EECF exhibited a significant (p <0.001) and optimal (100 mg/kg) gastroprotective effect by elevating SOD, CAT, and GSH levels, thereby minimizing the production of MDA and lowering the activity of MPO and vascular permeability. EECF also increased the rate of mucus production, decreased gastric motility, and completely suppressed the H+/K+ ATPase activity. Histopathological study confirmed the effectiveness of the extract in the prevention of peptic ulcer. The results obtained in this study demonstrated the gastro-protective effect of EECF via acidic antioxidant, anti-inflammatory, cytoprotective and anti-secretory mechanisms, which may justify its use as a substitute in peptic ulcer treatment.

Keywords: clematis flammula, superoxide dismutase, myeloperoxidase, ATPase, pump

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Authors: Taurino S. R. Neto, Epitácio D. S. Neto, Flávio Signorelli, Gustavo G. M. Balbi, Alex H. Higashi, Mário Luiz R. Monteiro, Eloisa Bonfá, Danieli C. O. Andrade, Leandro C. Zacharias

Abstract:

Purpose: To perform a multimodal evaluation, including the use of Optical Coherence Angiotomography (OCTA), in patients with primary antiphospholipid syndrome (PAPS) without ocular complaints and to compare them with healthy individuals. Methods: A complete structural and functional ophthalmological evaluation using OCTA and microperimetry (MP) exam in patients with PAPS, followed at a tertiary rheumatology outpatient clinic, was performed. All ophthalmologic manifestations were recorded and then statistical analysis was performed for comparative purposes; p <0.05 was considered statistically significant. Results: 104 eyes of 52 subjects (26 patients with PAPS without ocular complaints and 26 healthy individuals) were included. Among PAPS patients, 21 were female (80.8%) and 21 (80.8%) were Caucasians. Thrombotic PAPS was the main clinical criteria manifestation (100%); 65.4% had venous and 34.6% had arterial thrombosis. Obstetrical criteria were present in 34.6% of all thrombotic PAPS patients. Lupus anticoagulant was present in all patients. 19.2% of PAPS patients presented ophthalmologic findings against none of the healthy individuals. The most common retinal change was paracentral acute middle maculopathy (PAMM) (3 patients, 5 eyes), followed by drusen-like deposits (1 patient, 2 eyes) and pachychoroid pigment epitheliopathy (1 patient, 1 eye). Systemic hypertension and hyperlipidaemia were present in 100% of the PAPS patients with PAMM, while only six patients (26.1%) with PAPS without PAMM presented these two risk factors together. In the quantitative OCTA evaluation, we found significant differences between PAPS patients and controls in both the superficial vascular complex (SVC) and deep vascular complex (DVC) in the high-speed protocol, as well as in the SVC in the high-resolution protocol. In the analysis of the foveal avascular zone (FAZ) parameters, the PAPS group had a larger area of FAZ in the DVC using the high-speed method compared to the control group (p=0.047). In the quantitative analysis of the MP, the PAPS group had lower central (p=0.041) and global (p<0.001) retinal sensitivity compared to the control group, as well as in the sector analysis, with the exception of the inferior sector. In the quantitative evaluation of fixation stability, there was a trend towards worse stability in the PAPS subgroup with PAMM in both studied methods. Conclusions: PAMM was observed in 11.5% of PAPS patients with no previous ocular complaints. Systemic hypertension concomitant with hyperlipidemia was the most commonly associated risk factor for PAMM in patients with PAPS. PAPS patients present lower vascular density and retinal sensitivity compared to the control group, even in patients without PAMM.

Keywords: antiphospholipid syndrome, optical coherence angio tomography, optical coherence tomography, retina

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Authors: Atefeh Abedini, Fatemeh Razavi, Mihan Pourabdollah Toutkaboni, Hossein Mehravaran, Arda Kiani

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In countries with the highest prevalence of tuberculosis, such as Iran, the differentiation of malignant tumors from non-malignant is very important. In this study, which was conducted for the first time among the Iranian population, the utility of the ultrasonographic morphological characteristics in patients undergoing EBUS was used to distinguish the non-malignant versus malignant lymph nodes. The morphological characteristics of lymph nodes, which consist of size, shape, vascular pattern, echogenicity, margin, coagulation necrosis sign, calcification, and central hilar structure, were obtained during Endobronchial Ultrasound-Guided Trans-Bronchial Needle Aspiration and were compared with the final pathology results. During this study period, a total of 253 lymph nodes were evaluated in 93 cases. Round shape, non-hilar vascular pattern, heterogeneous echogenicity, hyperechogenicity, distinct margin, and the presence of necrosis sign were significantly higher in malignant nodes. On the other hand, the presence of calcification and also central hilar structure were significantly higher in the benign nodes (p-value ˂ 0.05). Multivariate logistic regression showed that size>1 cm, heterogeneous echogenicity, hyperechogenicity, the presence of necrosis signs and, the absence of central hilar structure are independent predictive factors for malignancy. The accuracy of each of the aforementioned factors is 42.29 %, 71.54 %, 71.90 %, 73.51 %, and 65.61 %, respectively. Of 74 malignant lymph nodes, 100% had at least one of these independent factors. According to our results, the morphological characteristics of lymph nodes based on Endobronchial Ultrasound-Guided Trans-Bronchial Needle Aspiration can play a role in the prediction of malignancy.

Keywords: EBUS-TBNA, malignancy, nodal characteristics, pathology

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Authors: Qing Zhang, Janak L. Pathak, Macro N. Helder, Richard T. Jaspers, Yin Xiao

Abstract:

Introduction: Nanomaterial-based bone regeneration is greatly influenced by the immune microenvironment. Tissue-engineered nanomaterials mediate the inflammatory response of macrophages to regulate bone regeneration. Silica nanoparticles have been widely used in tissue engineering-related preclinical studies. However, the effect of topological features on the surface of silica nanoparticles on the immune response of macrophages remains unknown. Purposes: The aims of this research are to compare the influences of normal and pollen-like silica nano-surface topography on macrophage immune responses and to obtain insight into their potential regulatory mechanisms. Method: Macrophages (RAW 264.7 cells) were exposed to mesoporous silica nanoparticles with normal morphology (MSNs) and pollen-like morphology (PMSNs). RNA-seq, RT-qPCR, and LSCM were used to assess the changes in expression levels of immune response-related genes and proteins. SEM and TEM were executed to evaluate the contact and adherence of silica nanoparticles by macrophages. For the assessment of the immunomodulation-mediated osteogenic potential, BMSCs were cultured with conditioned medium (CM) from LPS pre-stimulated macrophage cultures treated with MSNs or PMSNs. Osteoimmunomodulatory potential of MSNs and PMSNs in vivo was tested in a mouse cranial bone osteolysis model. Results: The results of the RNA-seq, RT-qPCR, and LSCM assays showed that PMSNs inhibited the expression of pro-inflammatory genes and proteins in macrophages. SEM images showed distinct macrophage membrane surface binding patterns of MSNs and PMSNs. MSNs were more evenly dispersed across the macrophage cell membrane, while PMSNs were aggregated. PMSNs-induced macrophage anti-inflammatory response was associated with upregulation of the cell surface receptor CD28 and inhibition of ERK phosphorylation. TEM images showed that both MSNs and PMSNs could be phagocytosed by macrophages, and inhibiting nanoparticle phagocytosis did not affect the expression of anti-inflammatory genes and proteins. Moreover, PMSNs-induced conditioned medium from macrophages enhanced BMP-2 expression and osteogenic differentiation mBMSCs. Similarly, PMSNs prevented LPS-induced bone resorption via downregulation of inflammatory reaction. Conclusions: PMSNs can promote bone regeneration by modulating osteoimmunological processes through surface topography. The study offers insights into how surface physical contact cues can modulate the regulation of osteoimmunology and provides a basis for the application of nanoparticles with pollen-like morphology to affect immunomodulation in bone tissue engineering and regeneration.

Keywords: physical contact, osteoimmunology, macrophages, silica nanoparticles, surface morphology, membrane receptor, osteogenesis, inflammation

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Authors: Spira A., Marabelle A., Kientop D., Moser E., Mumm J.

Abstract:

Background: Both interleukin-2 (IL-2) and interleukin-10 (IL-10) have been extensively studied for their stimulatory function on T cells and their potential to obtain sustainable tumor control in RCC, melanoma, lung, and pancreatic cancer as monotherapy, as well as combination with PD-1 blockers, radiation, and chemotherapy. While approved, IL-2 retains significant toxicity, preventing its widespread use. The significant efforts undertaken to uncouple IL-2 toxicity from its anti-tumor function have been unsuccessful, and early phase clinical safety observed with PEGylated IL-10 was not met in a blinded Phase 3 trial. Deka Biosciences has engineered a novel molecule coupling wild-type IL-2 to a high affinity variant of Epstein Barr Viral (EBV) IL-10 via a scaffold (scFv) that binds to epidermal growth factor receptors (EGFR). This patented molecule, termed DK210 (EGFR), is retained at high levels within the tumor microenvironment for days after dosing. In addition to overlapping and non-redundant anti-tumor function, IL-10 reduces IL-2 mediated cytokine release syndrome risks and inhibits IL-2 mediated T regulatory cell proliferation. Methods: DK210 (EGFR) is being evaluated in an open-label, dose-escalation (Phase 1) study with 5 (0.025-0.3 mg/kg) monotherapy dose levels and (expansion cohorts) in combination with PD-1 blockers, or radiation or chemotherapy in patients with advanced solid tumors overexpressing EGFR. Key eligibility criteria include 1) confirmed progressive disease on at least one line of systemic treatment, 2) EGFR overexpression or amplification documented in histology reports, 3) at least a 4 week or 5 half-lives window since last treatment, and 4) excluding subjects with long QT syndrome, multiple myeloma, multiple sclerosis, myasthenia gravis or uncontrolled infectious, psychiatric, neurologic, or cancer disease. Plasma and tissue samples will be investigated for pharmacodynamic and predictive biomarkers and genetic signatures associated with IFN-gamma secretion, aiming to select subjects for treatment in Phase 2. Conclusion: Through successful coupling of wild-type IL-2 with a high affinity IL-10 and targeting directly to the tumor microenvironment, DK210 (EGFR) has the potential to harness IL-2 and IL-10’s known anti-cancer promise while reducing immunogenicity and toxicity risks enabling safe concomitant cytokine treatment with other anti-cancer modalities.

Keywords: cytokine, EGFR over expression, interleukine-2, interleukine-10, clinical trial

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Authors: Delphine Morales, Florian Lombart, Agathe Truchot, Pauline Maire, Pascale Vigneron, Antoine Galmiche, Catherine Lok, Muriel Vayssade

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Targeted therapy molecules are used as a first-line treatment for metastatic melanoma with B-Raf mutation. Nevertheless, these molecules can cause side effects to patients and are efficient on 50 to 60 % of them. Indeed, melanoma cell sensitivity to targeted therapy molecules is dependent on tumor microenvironment (cell-cell and cell-extracellular matrix interactions). To better unravel factors modulating cell sensitivity to B-Raf inhibitor, we have developed and compared several melanoma models: from metastatic melanoma cells cultured as monolayer (2D) to a co-culture in a 3D dermal equivalent. Cell response was studied in different melanoma cell lines such as SK-MEL-28 (mutant B-Raf (V600E), sensitive to Vemurafenib), SK-MEL-3 (mutant B-Raf (V600E), resistant to Vemurafenib) and a primary culture of dermal human fibroblasts (HDFn). Assays have initially been performed in a monolayer cell culture (2D), then a second time on a 3D dermal equivalent (dermal human fibroblasts embedded in a collagen gel). All cell lines were treated with Vemurafenib (a B-Raf inhibitor) for 48 hours at various concentrations. Cell sensitivity to treatment was assessed under various aspects: Cell proliferation (cell counting, EdU incorporation, MTS assay), MAPK signaling pathway analysis (Western-Blotting), Apoptosis (TUNEL), Cytokine release (IL-6, IL-1α, HGF, TGF-β, TNF-α) upon Vemurafenib treatment (ELISA) and histology for 3D models. In 2D configuration, the inhibitory effect of Vemurafenib on cell proliferation was confirmed on SK-MEL-28 cells (IC50=0.5 µM), and not on the SK-MEL-3 cell line. No apoptotic signal was detected in SK-MEL-28-treated cells, suggesting a cytostatic effect of the Vemurafenib rather than a cytotoxic one. The inhibition of SK-MEL-28 cell proliferation upon treatment was correlated with a strong expression decrease of phosphorylated proteins involved in the MAPK pathway (ERK, MEK, and AKT/PKB). Vemurafenib (from 5 µM to 10 µM) also slowed down HDFn proliferation, whatever cell culture configuration (monolayer or 3D dermal equivalent). SK-MEL-28 cells cultured in the dermal equivalent were still sensitive to high Vemurafenib concentrations. To better characterize all cell population impacts (melanoma cells, dermal fibroblasts) on Vemurafenib efficacy, cytokine release is being studied in 2D and 3D models. We have successfully developed and validated a relevant 3D model, mimicking cutaneous metastatic melanoma and tumor microenvironment. This 3D melanoma model will become more complex by adding a third cell population, keratinocytes, allowing us to characterize the epidermis influence on the melanoma cell sensitivity to Vemurafenib. In the long run, the establishment of more relevant 3D melanoma models with patients’ cells might be useful for personalized therapy development. The authors would like to thank the Picardie region and the European Regional Development Fund (ERDF) 2014/2020 for the funding of this work and Oise committee of "La ligue contre le cancer".

Keywords: 3D human skin model, melanoma, tissue engineering, vemurafenib efficiency

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