Design, construction, sequence analysis and bioinformatics study of RTB-ipaD gene cassette: A new way in generation of Shigellosis vaccine

زمینه و هدف: شیگلا شایع ترین عامل اسهال می‌باشد. آنتی‌ژن پلاسمیدی IpaD برای تهاجم باکتری به درون سلول میزبان ضروری می‌باشد. یکی از چالش‌ها در باره واکسن مخاطی علیه شیگلا بر پایه پروتئین IpaD قدرت پایین آن می‌باشد. به نظر می‌رسد که با متصل کردن IpaD به یک ناقل و ادجوانت مناسب همچون زیر واحد B سم ریسین، می‌توان پروتئین IpaD را بسیار ایمنوژنیک نمود. این مطالعه به منظور تولید وکتور بیانی نوترکیب دارای کاست ژنی RTB-ipaD، آنالیز تعیین توالی و بررسی بیوانفورماتیکی آن انجام گرفته است. روش بررسی: در این مطالعه ژن های RTB و (163-483) ipaD در وکتور pGEM-T همسانه‌سازی شدند. ژن ipaD به روش برش آنزیمی با ژن RTB به همراه لینکر کد کننده GPGP در وکتور pGEM متصل شد. سپس قطعه کایمریک RTB-ipaD در وکتور بیانی pET28a(+) زیر همسانه‌سازی گردید. در پایان آنالیز تعیین توالی و بررسی بیوانفورماتیکی کاست ژنی انجام گرفت. یافته‌ها: صحت ساخت کاست ژنی RTB-ipaD در وکتور بیانی pET28a(+) با واکنش PCR و هضم آنزیمی مورد تایید قرار گرفت. نتایج حاصل از تعیین توالی کاست ژنی مطابق با توالی ذخیره شده در بانک ژنی بود. مطالعات بیوانفورماتیکی بر مبنای شاخص سازگاری کدون نشان داد این کاست قابلیت بیان در گیاه ترانس ژن و برخی سویه های باکتری اشرشیاکلی را دارد. نتیجه‌گیری: اتصال RTB به عنوان ناقل و ادجوانت به آنتی‌ژن IpaD رویکردی نوین و مطلوب در جهت تولید واکسن مخاطی شیگلوزیس می‌باش

DNA–Carbon Nanotube Complexation Affinity and Photoluminescence Modulation Are Independent

Short single-stranded DNA (ssDNA) has emerged as the natural polymer of choice for noncovalently functionalizing photoluminescent single-walled carbon nanotubes. In addition, specific empirically identified DNA sequences can be used to separate single species (chiralities) of nanotubes, with an exceptionally high purity. Currently, only limited general principles exist for designing DNA–nanotube hybrids amenable to separation processes, due in part to an incomplete understanding of the fundamental interactions between a DNA sequence and a specific nanotube structure, whereas even less is known in the design of nanotube-based sensors with determined optical properties. We therefore developed a combined experimental and analysis platform on the basis of time-resolved near-infrared fluorescence spectroscopy to extract the complete set of photoluminescence parameters that characterizes DNA–nanotube hybrids. Here, we systematically investigated the affinity of the d(GT)n oligonucleotide family for structurally defined carbon nanotubes by measuring photoluminescence response of the nanotube upon oligonucleotide displacement. We found, surprisingly, that the rate of displacement of the oligonucleotides is independent of the coverage on the nanotube, as inferred through the intrinsic optical properties of the hybrid. The kinetics of intensity modulation is essentially a single-exponential, and the time constants, which quantify the stability of DNA binding, span an order of magnitude. Surprisingly, these time constants do not depend on the intrinsic optical parameters within the hybrids, suggesting that the DNA–nanotube stability is not due to increased nanotube surface coverage by DNA. Further, a principal component analysis of the excitation and emission shifts along with intensity enhancement at equilibrium accurately identified the (8,6) nanotube as the partner chirality to (GT)6 ssDNA. When combined, the chirality-resolved equilibrium and kinetics data can guide the development of the DNA–nanotube pairs, with tunable stability and optical modulation. Additionally, this high-throughput optical platform could function as a primary screen for mapping the DNA-chirality recognition phase space

Carbon Nanotube–Liposome Complexes in Hydrogels for Controlled Drug Delivery via Near-Infrared Laser Stimulation

Externally controllable drug delivery systems are crucial for a variety of biological applications where the dosage and timing of drug delivery need to be adjusted based on disease diagnosis and progression. Here, we have developed an externally controllable drug delivery system by combining three extensively used platforms: hydrogels, liposomes, and single-walled carbon nanotubes (SWCNTs). We have developed carbon nanotube–liposome complexes (CLCs) and incorporated these structures into a 3D alginate hydrogel for use as an optically controlled drug delivery system. The CLC structures were characterized by using a variety of imaging and spectroscopic techniques, and an optimal SWCNT/lipid ratio was selected. The optimal CLCs were loaded with a model drug (FITC-Dex), incorporated into a hydrogel, and their release profile was studied. It was shown that release of the drug cargo can be triggered by using an NIR laser stimulation tuned to the optical resonance of a particular SWCNT species. It was further shown that the amount of released cargo can be tuned by varying the NIR stimulation time. This system demonstrates the externally controlled delivery of drug cargo and can be used for different applications including cancer chemotherapy delivery

Maximum Wireless Power Transmission Using Real-Time Single Iteration Adaptive Impedance Matching

Wireless power transfer (WPT) systems’ efficiency is significantly impacted by non-monotonic variations in the coupling coefficient. For very short distances or strong-coupling cases, the WPT efficiency is minimal at the natural resonant frequency, with two peaks around this frequency, known as the frequency splitting phenomenon. On the other hand, WPT capability decreases for long distances or weak coupling cases. Therefore, adaptive matching is required for WPT systems with varying distances, like wireless charging systems for electric vehicles (EVs). This paper first presents a detailed analysis of the frequency splitting phenomenon by studying the root locations of the WPT system’s transfer function. Then, a real-time fixed-frequency adaptive impedance matching (IM) method is proposed, in which the amplitude and phase of the input impedance is estimated using the average active power, the average reactive power, and the amplitude of input voltage. Unlike traditional search-and-find techniques, the proposed method calculates the optimal IM network parameters only in a single iteration, which improves the convergent speed. A scaled-down 20-Watt prototype controlled by the TMSF2812 is fabricated and used to validate the effectiveness of the proposed method over a wide range of coil-to-coil distances

Effect of the exercise programme on the quality of life of prostate cancer survivors: A randomized controlled trial

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Climate change adaptation in conflict-affected countries:A systematic assessment of evidence

People affected by conflict are particularly vulnerable to climate shocks and climate change, yet little is known about climate change adaptation in fragile contexts. While climate events are one of the many contributing drivers of conflict, feedback from conflict increases vulnerability, thereby creating conditions for a vicious cycle of conflict. In this study, we carry out a systematic review of peer-reviewed literature, taking from the Global Adaptation Mapping Initiative (GAMI) dataset to documenting climate change adaptation occurring in 15 conflict-affected countries and compare the findings with records of climate adaptation finance flows and climate-related disasters in each country. Academic literature is sparse for most conflict-affected countries, and available studies tend to have a narrow focus, particularly on agriculture-related adaptation in rural contexts and adaptation by low-income actors. In contrast, multilateral and bilateral funding for climate change adaptation addresses a greater diversity of adaptation needs, including water systems, humanitarian programming, and urban areas. Even among the conflict-affected countries selected, we find disparity, with several countries being the focus of substantial research and funding, and others seeing little to none. Results indicate that people in conflict-affected contexts are adapting to climate change, but there is a pressing need for diverse scholarship across various sectors that documents a broader range of adaptation types and their results

Reprogramming the assembly of unmodified DNA with a small molecule

The ability of DNA to store and encode information arises from base pairing of the four-letter nucleobase code to form a double helix. Expanding this DNA ‘alphabet’ by synthetic incorporation of new bases can introduce new functionalities and enable the formation of novel nucleic acid structures. However, reprogramming the self-assembly of existing nucleobases presents an alternative route to expand the structural space and functionality of nucleic acids. Here we report the discovery that a small molecule, cyanuric acid, with three thymine-like faces reprogrammes the assembly of unmodified poly(adenine) (poly(A)) into stable, long and abundant fibres with a unique internal structure. Poly(A) DNA, RNA and peptide nucleic acid all form these assemblies. Our studies are consistent with the association of adenine and cyanuric acid units into a hexameric rosette, which brings together poly(A) triplexes with a subsequent cooperative polymerization. Fundamentally, this study shows that small hydrogen-bonding molecules can be used to induce the assembly of nucleic acids in water, which leads to new structures from inexpensive and readily available materials

Kinematic analysis of the daily activity of drinking from a glass in a population with cervical spinal cord injury

Background Three-dimensional kinematic analysis equipment is a valuable instrument for studying the execution of movement during functional activities of the upper limbs. The aim of this study was to analyze the kinematic differences in the execution of a daily activity such as drinking from a glass between two groups of patients with tetraplegia and a control group. Methods A total of 24 people were separated into three groups for analysis: 8 subjects with metameric level C6 tetraplegia, 8 subjects with metameric level C7 tetraplegia and 8 control subjects (CG). A set of active markers that emit infrared light were positioned on the upper limb. Two scanning units were used to record the sessions. The activity of drinking from a glass was broken down into a series of clearly identifiable phases to facilitate analysis. Movement times, velocities, and the joint angles of the shoulder, elbow and wrist in the three spatial planes were the variables analyzed. Results The most relevant differences between the three groups were in the wrist. Wrist palmar flexion during the back transport phase was greater in the patients with C6 and C7 tetraplegia than in the CG, whereas the highest wrist dorsal flexion values were in forward transport in the subjects with C6 or C7 tetraplegia, who required complete activation of the tenodesis effect to complete grasping. Conclusions A detailed description was made of the three-dimensional kinematic analysis of the task of drinking from a glass in healthy subjects and in two groups of patients with tetraplegia. This was a useful application of kinematic analysis of upper limb movement in a clinical setting. Better knowledge of the execution of this movement in each of these groups allows therapeutic recommendations to be specifically adapted to the functional deficit present. This information can be useful in designing wearable robots to compensate the performance of AVD, such as drinking, in people with cervical SCI

Monitoring of Regulatory T Cell Frequencies and Expression of CTLA-4 on T Cells, before and after DC Vaccination, Can Predict Survival in GBM Patients

PURPOSE: Dendritic cell (DC) vaccines have recently emerged as an innovative therapeutic option for glioblastoma patients. To identify novel surrogates of anti-tumor immune responsiveness, we studied the dynamic expression of activation and inhibitory markers on peripheral blood lymphocyte (PBL) subsets in glioblastoma patients treated with DC vaccination at UCLA. EXPERIMENTAL DESIGN: Pre-treatment and post-treatment PBL from 24 patients enrolled in two Phase I clinical trials of dendritic cell immunotherapy were stained and analyzed using flow cytometry. A univariate Cox proportional hazards model was utilized to investigate the association between continuous immune monitoring variables and survival. Finally, the immune monitoring variables were dichotomized and a recursive partitioning survival tree was built to obtain cut-off values predictive of survival. RESULTS: The change in regulatory T cell (CD3(+)CD4(+)CD25(+)CD127(low)) frequency in PBL was significantly associated with survival (p = 0.0228; hazard ratio = 3.623) after DC vaccination. Furthermore, the dynamic expression of the negative co-stimulatory molecule, CTLA-4, was also significantly associated with survival on CD3(+)CD4(+) T cells (p = 0.0191; hazard ratio = 2.840) and CD3(+)CD8(+) T cells (p = 0.0273; hazard ratio = 2.690), while that of activation markers (CD25, CD69) was not. Finally, a recursive partitioning tree algorithm was utilized to dichotomize the post/pre fold change immune monitoring variables. The resultant cut-off values from these immune monitoring variables could effectively segregate these patients into groups with significantly different overall survival curves. CONCLUSIONS: Our results suggest that monitoring the change in regulatory T cell frequencies and dynamic expression of the negative co-stimulatory molecules on peripheral blood T cells, before and after DC vaccination, may predict survival. The cut-off point generated from these data can be utilized in future prospective immunotherapy trials to further evaluate its predictive validity