Oxygen vacancy-rich Fe2(MoO4)3 combined with MWCNTs for electrochemical sensors of fentanyl and its analogs.

Hundreds of thousands of people dying from the abuse of fentanyl and its analogs. Hence, the development of an efficient and highly accurate detection method is extremely relevant and challenging. Therefore, we proposed the introduction of oxygen defects into Fe2(MoO4)3 nanoparticles for improving the catalyst performance and combining it with multi-walled carbon nanotubes (MWCNTs) for electrochemical detection of fentanyl and its analogs. Oxygen vacancy-rich Fe2(MoO4)3 (called r-Fe2(MoO4)3) nanoparticles were successfully synthesized and characterized in detail by scanning electron microscopy (SEM), transmission electron microscopy (TEM), energy-dispersive spectrometry (EDS), X-ray diffraction (XRD), Fourier transform infrared (FT-IR), Raman spectra, BET, X-ray photoelectron spectroscopy (XPS), and electron paramagnetic resonance (EPR) and investigated by comparison with oxygen vacancy-poor Fe2(MoO4)3 (called p-Fe2(MoO4)3). The obtained oxygen vacancy-rich Fe2(MoO4)3 was ultrasonically composited with MWCNTs for modification of glassy carbon electrodes (GCEs) used for the electrochemical detection of fentanyl and its analogs. The modified MWCNT-GCE showed ultrasensitivity to fentanyl, sufentanil, alfentanil, and acetylfentanyl with limits of detection (LOD) of 0.006 µmol·L-1, 0.008 µmol·L-1, 0.018 µmol·L-1, and 0.024 µmol·L-1, respectively, and could distinguish among the four drugs based on their peak voltages. Besides, the obtained r-Fe2(MoO4)3/MWCNT composite also exhibited high repeatability, selectivity, and stability. It showed satisfactory detection performance on real samples, with recoveries of 70.53 ~ 94.85% and 50.98 ~ 82.54% in serum and urine for the four drugs in a concentration range 0.2 ~ 1 µM, respectively. The experimental results confirm that the introduction of oxygen vacancies effectively improves the sensitivity of fentanyl electrochemical detection, and this work provides some inspiration for the development of catalytic materials for electrochemical sensors with higher sensitivity.

Insights into the solvation chemistry in liquid electrolytes for lithium-based rechargeable batteries.

Lithium-based rechargeable batteries have dominated the energy storage field and attracted considerable research interest due to their excellent electrochemical performance. As indispensable and ubiquitous components, electrolytes play a pivotal role in not only transporting lithium ions, but also expanding the electrochemical stable potential window, suppressing the side reactions, and manipulating the redox mechanism, all of which are closely associated with the behavior of solvation chemistry in electrolytes. Thus, comprehensively understanding the solvation chemistry in electrolytes is of significant importance. Here we critically reviewed the development of electrolytes in various lithium-based rechargeable batteries including lithium-metal batteries (LMBs), nonaqueous lithium-ion batteries (LIBs), lithium-sulfur batteries (LSBs), lithium-oxygen batteries (LOBs), and aqueous lithium-ion batteries (ALIBs), and emphasized the effects of interactions between cations, anions, and solvents on solvation chemistry, and functions of solvation chemistry in different types of electrolytes (strong solvating electrolytes, moderate solvating electrolytes, and weak solvating electrolytes) on the electrochemical performance and redox mechanism in the abovementioned rechargeable batteries. Specifically, the significant effects of solvation chemistry on the stability of electrode-electrolyte interphases, suppression of lithium dendrites in LMBs, inhibition of the co-intercalation of solvents in LIBs, improvement of anodic stability at high cut-off voltages in LMBs, LIBs and ALIBs, regulation of redox pathways in LSBs and LOBs, and inhibition of hydrogen/oxygen evolution reactions in LOBs are thoroughly summarized. Finally, the review concludes with a prospective outlook, where practical issues of electrolytes, advanced in situ/operando techniques to illustrate the mechanism of solvation chemistry, and advanced theoretical calculation and simulation techniques such as "material knowledge informed machine learning" and "artificial intelligence (AI) + big data" driven strategies for high-performance electrolytes have been proposed.

Application of immersive virtual reality (IVR) for the care of common diseases of older adults course: A mixed methods study.

OBJECTIVE: This study aimed to enhance understanding, engagement, and learning efficiency in the course "The Care of Common Diseases of Older Adults" using a developed Immersive Virtual Reality(IVR) system. METHODS: A mixed-methods study with 32 students was conducted. The quantitative part involved a randomized controlled trial, and the qualitative part included thematic interviews with students and teachers. RESULTS: The intervention group using the IVR system showed significant improvements in positivity and performance evaluation scores (P < 0.05) compared to the control group. Negative affect scores also decreased significantly (P < 0.05). Qualitative data from interviews supported the quantitative findings, highlighting increased curiosity, learning enthusiasm, and academic performance. CONCLUSION: IVR significantly enhances learning by stimulating curiosity and active participation, making education more accessible and improving student performance. Future IVR enhancements should focus on user-friendliness and empathetic feedback in adult care.

Interface Engineering via Manipulating Solvation Chemistry for Liquid Lithium-Ion Batteries Operated ≥ 100 °C.

High-performance and temperature-resistant lithium-ion batteries (LIBs), which are able to operate at elevated temperatures (i.e., >60 °C) are highly demanded in various fields, especially in military or aerospace exploration. However, their applications were  impeded by the poor electrochemical performance and unsatisfying safety issues, which was induced by the severe side reactions between electrolytes and electrodes at high temperatures. Herein, with the synergetic effects of solvation chemistry and functional additive in the elaborately designed weakly solvating electrolyte, a unique robust organic/inorganic hetero-interphase, composed of gradient F, B-rich inorganic components and homogeneously distributed Si-rich organic components, was successfully constructed on both cathodes and anodes, which would effectively inhibit the constant decomposition of electrolytes and dissolution of transition metal ions. As a result, both cathodes and anodes, without compromising their low-temperature performance, operate at temperatures ≥100 ℃, with excellent capacity retentions of 96.1 % after 500 cycles and 93.5% after ≥200 cycles, respectively, at 80 ℃. Ah-level LiCoO2||graphite full cells with a cut-off voltage of 4.3 V also exhibited superior temperature-resistance with a capacity retention of 89.9% at temperature as high as 120 ℃. Moreover, the fully charged pouch cells exhibited highly enhanced safety, demonstrating their potentials in practical applications at ultrahigh temperatures.

Chitosan Sponge/Cu-WO3-x Composite for Photodynamic Therapy of Wound Infection.

One of the potential treatments for bacterial wound infections is photodynamic therapy. WO3-x semiconductor materials can generate reactive oxygen species when exposed to light, which can inactivate bacteria. In this work, we improved their photocatalytic performance by doping WO3-x with Cu. The wound dressing was prepared by loading Cu-WO3-x into a highly biocompatible chitosan sponge. The composite sponge dressing showed significant inactivation of Escherichia coli and Staphylococcus aureus, and in vitro toxicity assays on L929 cells demonstrated the biosafety of the dressing. Through in vivo wound healing trials, composite sponge dressings have been shown to accelerate wound healing, and this composite chitosan sponge can be possibly used for photodynamic therapy of bacterial wound infections.

PVA/WO3-x/Ag2WO4 Hydrogel for Enhanced Photocatalytic Disinfection and Accelerated Wound Healing.

Bacterial wound infections are one of the growing health and safety threats to the public. In this study, WO3-x/Ag2WO4 photocatalysts were synthesized, and heterogeneous structures were constructed for non-antibiotic bactericidal use. Due to the heterostructure constructed with Ag2WO4, the photogenerated carrier separation efficiency and reactive oxygen generation capacity of WO3-x were improved, which in turn improved the inactivation rate of bacteria. Also, this photocatalyst was loaded into PVA hydrogel for photodynamic treatment of bacterial wound infections. This hydrogel dressing was demonstrated to have good biosafety by in vitro cytotoxicity tests and to have a wound healing-promoting effect by in vivo wound healing experiments. This light-driven antimicrobial hydrogel has the potential ability to treat bacterial wound infections.

A series of ultrasensitive electrocatalysts Fe-MOF/MWCNTs for fentanyl determination.

Electrochemical determination of synthetic opioids such as fentanyl is meaningful but still challenging no matter from a social or academic perspective. Herein, we report a series of novel electrocatalysts based on Fe-containing metal-organic frameworks and multi-walled carbon nanotubes (Fe-MOF/MWCNTs). The obtained Fe-MOF/MWCNT electrode materials all show ultrasensitivity on fentanyl determination. In particular, MOF-235/MWCNTs even exhibit an ultra-low limit of detection (LOD) of 0.03 µM with a wide linear range from 0.1 to 50 µM. Besides, this series of Fe-MOF/MWCNTs also displays excellent repeatability, selectivity, and stability. Moreover, they show good performance in real sample detection and achieve good recovery of 95.47%-102.41% and 96.62%-103.15% in blood and urine samples, respectively. This high performance in fentanyl determination is mainly contributed by the Fenton-like process and the adsorption function of the Fe-MOF. Therefore, these novel Fe-MOF/MWCNTs are promising electrocatalysts for point-of-care device fabrication and also have potential applications in fentanyl rapid test technology.

Generation of vector vortex beams by axially symmetric sheared polymer network liquid crystals.

Liquid crystals have been widely used in optoelectronic devices because of their fast response and excellent electro-optic properties. Featuring a unique ability to manipulate light, they are also proposed as a good candidate in topological photonics for further applications. In this study, an axially symmetric sheared polymer network liquid crystal (ASPNLC) is fabricated to demonstrate vector vortex beams. Linearly and circularly polarized light is used to illuminate the sample, and the output vector vortex beams generated from the ASPNLC indicate that the polarization states of the output beams are dependent on the polarization of the incident light. The measured phenomena are modeled on the bases of phase retardation and Jones calculus to eventually calculate the polarization-resolved intensity profiles accordingly. Hence, our experimental study provides a holistic understanding of the method for generating vector vortex beams by an ASPNLC, which is expected to enhance the knowledge of optical mechanisms for liquid crystal applications.

Highly sensitive RI and temperature sensor based on an asymmetric fiber coupler.

We propose and demonstrate a highly sensitive refractive index (RI) and temperature sensor based on an asymmetric fiber coupler (AFC). The AFC was fabricated by weak fusion of a pre-stretched single-mode fiber and a few-mode fiber. An ultra-sensitivity RI can be achieved near the dispersion turning point (DTP). The proposed RI sensor achieves a high RI sensitivity of -10,662.4nm/RIU within the RI range of 1.31-1.35. By packaging the AFC into polydimethylsiloxane (PDMS), the temperature sensitivity reaches 11.44 nm/°C. The proposed AFC with high RI and temperature sensitivity can be potentially used in the field of chemical monitoring, biochemical detection, and clinical diagnosis.

Prognostic value of quantitative flow ratio measured immediately after drug-coated balloon angioplasty for in-stent restenosis.

OBJECTIVES: This study aimed to evaluate prognostic value of quantitative flow ratio (QFR) in drug-coated balloon (DCB) angioplasty for in-stent restenosis (ISR). BACKGROUND: There is a high incidence of recurrent ISR after DCB angioplasty. QFR is a novel method for fast computation of fractional flow reserve for the target vessel based on quantitative coronary angiography (QCA) and fluid dynamics algorithms. METHODS: Patients participating in the RESTORE ISR China randomized trial were enrolled and classified into the recurrent restenosis group and the non-recurrent restenosis group. The binary classifications followed the QCA standards of ISR. Clinical and angiographic characteristics of the groups were analyzed, and the QFRs before and after lesion preparation and after final DCB angioplasty were measured and compared. RESULTS: A total of 208 patients who underwent follow-up angiography were enrolled in the study, with 226 lesions measured in total. QFR value after DCB angioplasty (odds ratio [OR] 0.88; 95% confidence interval [CI] 0.83-0.93; p < .0001 for 1 mm increase), lesion length (OR: 1.08; 95% CI: 1.01-1.15; p = .017), and vessel caliber lumen diameter (OR: 0.35; 95% CI 0.13-0.89; p = .027) were independently associated with recurrent restenosis after DCB angioplasty. The optimal QFR cut-off value was determined to be 0.90 with a sensitivity of 0.94, specificity of 0.56, and accuracy of 0.79 in predicting recurrent restenosis. CONCLUSIONS: The QFR value after DCB angioplasty is a promising predictor of DES ISR.

In Vitro and In Vivo Comparative Evaluation of a Shellac-Ammonium Paclitaxel-Coated Balloon versus a Benchmark Device.

OBJECTIVES: The present study was designed to compare the characteristics and performance regarding drug delivery of a novel drug-coated balloon (DCB) to a benchmark device (Restore® versus SeQuent® Please) in an in vitro and in vivo model. BACKGROUND: Although Restore® and SeQuent® are both paclitaxel-coated, they use different coating excipient, shellac-ammonium salt and iopromide, respectively. Preclinical study comparing these two different commercial DCBs regarding their characteristics and effects on early vascular response is sparse. METHODS: Restore® and SeQuent® DCBs were scanned with electron microscopy for surface characteristic assessment. Both DCBs were transported in an in vitro vessel model for the evaluation of drug wash-off rate and particulate formation. Eighteen coronary angioplasties with either Restore® or SeQuent® DCBs were conducted in 6 swine (three coronary vessels each). Histopathological images of each vessel were evaluated for vessel injury. RESULTS: The surface of Restore® DCB was smooth and evenly distributed with hardly visible crystal, while SeQuent® DCB showed a rougher surface with relatively larger apparent crystals. Restore® DCB had a lower drug wash-off rate and fewer large visible particles, compared to the SeQuent® DCB. No significant difference in mean injure score was found between Restore® and SeQuent® group. CONCLUSION: Our results suggest that Restore® is better in preclinical performance regarding less release of particles and lower drug wash-off rate as compared to SeQuent® Please. The Restore® DCB, using stable amorphous coating and shellac-ammonium salt as an excipient, appears to provide an advantage in drug delivery efficacy; however, further clinical studies are warranted.

Exosomal lncRNA UCA1 from cancer-associated fibroblasts enhances chemoresistance in vulvar squamous cell carcinoma cells.

AIM: In the current work, we aimed to explore whether Cancer-associated fibroblasts (CAF) exosomes played crucial roles in vulvar squamous cell carcinoma (VSCC) chemoresistance via mediating long noncoding RNAs (lncRNA). METHODS: The IC50 value and cell apoptosis were assessed by the Cell Counting-8 Kit (CCK-8) assay and flow cytometry, respectively. Western blot analysis was used for the measurement of protein levels. The levels of urothelial cancer-associated 1 (UCA1), miR-103a and WEE1 G2 checkpoint kinase (WEE1) mRNA were detected by quantitative real-time polymerase chain reaction (qRT-PCR). The target relationships among miR-103a, UCA1 and WEE1 were confirmed by dual-luciferase reporter assays. Xenograft model mice were established to observe the impact of exosomal UCA1 on cisplatin (CDDP) resistance in vivo. RESULTS: Our data indicated that CAF enhanced CDDP resistance of VSCC cells in vitro. Extracellular UCA1 was transferred by exosomes derived from CAF. Exosomal UCA1 derived from CAF conferred VSCC cell resistance to CDDP. Moreover, UCA1 functioned as a miR-103a sponge in VSCC cells. The promotion of exosomal UCA1 on VSCC cell resistance to CDDP was mediated by miR-103a. WEE1 was a direct target of miR-103a, and exosomal miR-103a from CAF weakened CDDP resistance of VSCC cells by WEE1. Furthermore, exosomal UCA1 regulated WEE1 expression through sponging miR-103a. Additionally, exosomal UCA1 enhanced tumor growth and CDDP resistance in vivo. CONCLUSION: Our findings suggested exosomal UCA1 derived from CAF conferred VSCC cell resistance to CDDP in vitro and in vivo at least partly through the miR-103a/WEE1 axis, highlighting a novel therapeutic method for improving the clinical benefits of CDDP chemotherapy in VSCC patients.

Optimization-Based Approaches for Minimizing Deployment Costs for Wireless Sensor Networks with Bounded Estimation Errors.

As wireless sensor networks have become more prevalent, data from sensors in daily life are constantly being recorded. Due to cost or energy consumption considerations, optimization-based approaches are proposed to reduce deployed sensors and yield results within the error tolerance. The correlation-aware method is also designed in a mathematical model that combines theoretical and practical perspectives. The sensor deployment strategies, including XGBoost, Pearson correlation, and Lagrangian Relaxation (LR), are determined to minimize deployment costs while maintaining estimation errors below a given threshold. Moreover, the results significantly ensure the accuracy of the gathered information while minimizing the cost of deployment and maximizing the lifetime of the WSN. Furthermore, the proposed solution can be readily applied to sensor distribution problems in various fields.

An Optimization-Based Orchestrator for Resource Access and Operation Management in Sliced 5G Core Networks.

Network slicing is a promising technology that network operators can deploy the services by slices with heterogeneous quality of service (QoS) requirements. However, an orchestrator for network operation with efficient slice resource provisioning algorithms is essential. This work stands on Internet service provider (ISP) to design an orchestrator analyzing the critical influencing factors, namely access control, scheduling, and resource migration, to systematically evolve a sustainable network. The scalability and flexibility of resources are jointly considered. The resource management problem is formulated as a mixed-integer programming (MIP) problem. A solution approach based on Lagrangian relaxation (LR) is proposed for the orchestrator to make decisions to satisfy the high QoS applications. It can investigate the resources required for access control within a cost-efficient resource pool and consider allocating or migrating resources efficiently in each network slice. For high system utilization, the proposed mechanisms are modeled in a pay-as-you-go manner. Furthermore, the experiment results show that the proposed strategies perform the near-optimal system revenue to meet the QoS requirement by making decisions.

Optimization-Based Resource Management Algorithms with Considerations of Client Satisfaction and High Availability in Elastic 5G Network Slices.

A combined edge and core cloud computing environment is a novel solution in 5G network slices. The clients' high availability requirement is a challenge because it limits the possible admission control in front of the edge cloud. This work proposes an orchestrator with a mathematical programming model in a global viewpoint to solve resource management problems and satisfying the clients' high availability requirements. The proposed Lagrangian relaxation-based approach is adopted to solve the problems at a near-optimal level for increasing the system revenue. A promising and straightforward resource management approach and several experimental cases are used to evaluate the efficiency and effectiveness. Preliminary results are presented as performance evaluations to verify the proposed approach's suitability for edge and core cloud computing environments. The proposed orchestrator significantly enables the network slicing services and efficiently enhances the clients' satisfaction of high availability.

Prognostic roles of time to positivity of blood cultures in patients with Escherichia coli bacteremia.

The time to positivity (TTP) of blood cultures has been considered a predictor of clinical outcomes for bacteremia. This retrospective study aimed to determine the clinical value of TTP for the prognostic assessment of patients with Escherichia coli bacteremia. A total of 167 adult patients with E.coli bacteremia identified over a 22-month period in a 3500-bed university teaching hospital in China were studied. The standard cut-off TTP was 11 h in the patient cohort. The septic shock occurred in 27.9% of patients with early TTP (⩽11 h) and in 7.1% of those with a prolonged TTP (>11 h) (P = 0.003). The mortality rate was significantly higher for patients in the early than in the late group (17.7% vs. 4.0%, P < 0.001). Multivariate analysis showed that an early TTP (OR 4.50, 95% CI 1.70-11.93), intensive care unit admission (OR 8.39, 95% CI 2.01-35.14) and neutropenia (OR 4.20, 95% CI 1.55-11.40) were independently associated with septic shock. Likewise, the independent risk factors for mortality of patients were an early TTP (OR 3.80, 95% CI 1.04-12.90), intensive care unit admission (OR 6.45; 95% CI 1.14-36.53), a Pittsburgh bacteremia score ⩾2 (OR 4.34, 95% CI 1.22-15.47) and a Charlson Comorbidity Index ⩾3 (OR 11.29, 95% CI 2.81-45.39). Overall, a TTP for blood cultures within 11 h appears to be associated with worse outcomes for patients with E.coli bacteremia.

Broadband tunable single-longitudinal-mode erbium-doped fiber ring laser based on a microfiber knot resonator.

We propose and demonstrate a broadband tunable single-longitudinal-mode (SLM) erbium-doped fiber laser based on a microfiber knot resonator (MKR). The MKR is made from a double-ended fiber taper and employed for SLM filtering based on the Vernier effect. An unpumped erbium-doped fiber is used in the fiber laser cavity to suppress mode-hopping for a stabilized SLM laser operation. When combined with an optical fiber filter, widely tunable SLM laser generation is achieved. The proposed SLM laser can be tuned from 1545 to 1565 nm with a high side-mode suppression ratio of about 55 dB and a high stability.

PAK4, a target of miR-9-5p, promotes cell proliferation and inhibits apoptosis in colorectal cancer.

BACKGROUND: Colorectal cancer (CRC) is a leading cause of cancer-related death worldwide. P21-activated kinase 4 (PAK4) and miR-9-5p have emerged as attractive therapeutic targets in several tumor types, but in CRC, the regulation of their biological function and their target association remain unclear. METHODS: The expression of PAK4 in CRC tissues was determined using quantitative real-time PCR and immunohistochemistry analyses. The targeted regulation between miR-9-5p and PAK4 was predicted and confirmed with bioinformatics analysis and the dual-luciferase reporter assay. Functional experiments, including the MTT assay and flow cytometry, were performed to investigate the impact of PAK4 knockdown and miR-9-5p overexpression on cell proliferation and apoptosis in CRC cells. RESULTS: We found that the expression of PAK4 was upregulated in CRC tissues. PAK4 knockdown significantly suppressed cell proliferation and promoted apoptosis in cells of the CRC cell lines HCT116 and SW1116. We also found that miR-9-5p directly targeted the 3'-UTR of PAK4 mRNA and negatively regulated its expression. The degree of downregulation of miR-9-5p inversely correlated with PAK4 expression. Intriguingly, enforced expression of miR-9-5p suppressed cell proliferation and promoted apoptosis. This could be partially reversed by PAK4 overexpression. CONCLUSION: These results suggest that miR-9-5p targeting of PAK4 could have therapeutic potential for CRC treatment.

Temporal-Spatial Establishment of Initial Niche for the Primary Spermatogonial Stem Cell Formation Is Determined by an ARID4B Regulatory Network.

During neonatal testis development, centrally located gonocytes migrate to basement membrane of the seminiferous cords, where physical contact with a niche established by Sertoli cells is essential for transition of gonocytes into spermatogonial stem cells (SSCs). To provide structural support and signaling stimuli for the gonocyte-to-SSC transition that occurs at a specific location during a finite phase, temporal-spatial establishment of the niche is critical. To date, the factors that guide Sertoli cells to establish the initial stem cell niche remain largely unknown. Using the Sertoli cell-specific Arid4b knockout (Arid4bSCKO) mice, we demonstrated that ablation of AT-rich interaction domain 4B (ARID4B) resulted in abnormal detachment of Sertoli cells from the basement membrane of seminiferous cords during the gonocyte-to-SSC transition phase, suggesting failure to establish a niche for the SSC formation. Without support by a niche environment, gonocytes showed disarranged cell distribution in the Arid4bSCKO testes and underwent apoptosis. The commitment of gonocytes to differentiate into the spermatogonial lineage was broken and the capability of SSCs to self-renew and differentiate was also impaired. Gene expression profiling revealed the molecular mechanisms responsible for the phenotypic changes in the Arid4bSCKO testes, by identifying genes important for stem cell niche function as downstream effectors of ARID4B, including genes that encode gap junction protein alpha-1, KIT ligand, anti-Müllerian hormone, Glial cell-line derived neurotrophic factor, inhibin alpha, inhibin beta, and cytochrome P450 family 26 subfamily b polypeptide 1. Our results identified ARID4B as a master regulator of a signaling network that governs the establishment of a niche during the critical gonocyte-to-SSC transition phase to control the fate of gonocytes and SSCs. Stem Cells 2017;35:1554-1565.

Structure-based design of free fatty acid receptor 1 agonists bearing non-biphenyl scaffold.

The free fatty acid receptor 1 (FFA1) enhances the glucose-stimulated insulin secretion without the risk of hypoglycemia. However, most of FFA1 agonists have a common biphenyl moiety, leading to a relative deprivation in structure types. Herein, we describe the exploration of non-biphenyl scaffold based on the co-crystal structure of FFA1 to increase additional interactions with the lateral residues, which led to the identification of lead compounds 3 and 9. In induced-fit docking study, compound 3 forms an edge-on interaction with Trp150 by slightly rotating the indole ring of Trp150, and compound 9 has additional hydrogen bond and δ-π interactions with Leu135, which demonstrated the feasibility of our design strategy. Moreover, lead compounds 3 and 9 revealed improved polar surface area compared to GW9508, and have considerable hypoglycemic effects in mice. This structure-based study might inspire us to design more promising FFA1 agonists by increasing additional interactions with the residues outside of binding pocket.