Antiviral peptides inhibiting the main protease of SARS-CoV-2 investigated by computational screening and in vitro protease assay.

The main protease (Mpro) of severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) plays an important role in viral replication and transcription and received great attention as a vital target for drug/peptide development. Therapeutic agents such as small-molecule drugs or peptides that interact with the Cys-His present in the catalytic site of Mpro are an efficient way to inhibit the protease. Although several emergency-approved vaccines showed good efficacy and drastically dropped the infection rate, evolving variants are still infecting and killing millions of people globally. While a small-molecule drug (Paxlovid) received emergency approval, small-molecule drugs have low target specificity and higher toxicity. Besides small-molecule drugs, peptide therapeutics are thus gaining increasing popularity as they are easy to synthesize and highly selective and have limited side effects. In this study, we investigated the therapeutic value of 67 peptides targeting Mpro using molecular docking. Subsequently, molecular dynamics (MD) simulations were implemented on eight protein-peptide complexes to obtain molecular-level information on the interaction between these peptides and the Mpro active site, which revealed that temporin L, indolicidin, and lymphocytic choriomeningitis virus (LCMV) GP1 are the best candidates in terms of stability, interaction, and structural compactness. These peptides were synthesized using the solid-phase peptide synthesis protocol, purified by reversed-phase high-performance liquid chromatography (RP-HPLC), and authenticated by mass spectrometry (MS). The in vitro fluorometric Mpro activity assay was used to validate the computational results, where temporin L and indolicidin were observed to be very active against SARS-CoV-2 Mpro with IC50 values of 38.80 and 87.23 µM, respectively. A liquid chromatography-MS (LC-MS) assay was developed, and the IC50 value of temporin L was measured at 23.8 µM. The solution-state nuclear magnetic resonance (NMR) structure of temporin L was determined in the absence of sodium dodecyl sulfate (SDS) micelles and was compared to previous temporin structures. This combined investigation provides critical insights and assists us to further develop peptide inhibitors of SARS-CoV-2 Mpro through structural guided investigation.

Bilateral Squatting Mechanics Are Associated With Landing Mechanics in Anterior Cruciate Ligament Reconstruction Patients.

BACKGROUND: Proper lower extremity biomechanics during bilateral landing is important for reducing injury risk in athletes returning to sports after anterior cruciate ligament reconstruction (ACLR). Although landing is a quick ballistic movement that is difficult to modify, squatting is a slower cyclic movement that is ideal for motor learning. HYPOTHESIS: There is a relationship between lower extremity biomechanics during bilateral landing and bilateral squatting in patients with an ACLR. STUDY DESIGN: Descriptive laboratory study. METHODS: A total of 41 patients after a unilateral ACLR (24 men, 17 women; 5.9 ± 1.4 months after ACLR) completed 15 unweighted bilateral squats and 10 bilateral stop-jumps. Three-dimensional lower extremity kinematics and kinetics were collected, and peak knee abduction angle, knee abduction/adduction range of motion, peak vertical ground-reaction force limb symmetry index (LSI), vertical ground-reaction force impulse LSI, and peak knee extension moment LSI were computed during the descending phase of the squatting and landing tasks. Wilcoxon signed-rank tests were used to compare each outcome between limbs, and Spearman correlations were used to compare outcomes between the squatting and landing tasks. RESULTS: The peak vertical ground reaction force, the vertical ground reaction force impulse, and the peak knee extension moment were reduced in the surgical (Sx) limb relative to the nonsurgical (NSx) limb during both the squatting and landing tasks (P < .001). The relationship between squatting and landing tasks was strong for the peak knee abduction angle (R = 0.697-0.737; P < .001); moderate for the frontal plane knee range of motion (NSx: R = 0.366, P = .019; Sx: R = 0.418, P = 0.007), the peak knee extension moment LSI (R = 0.573; P < .001), the vertical ground reaction force impulse LSI (R = 0.382; P < .014); and weak for the peak vertical ground reaction force LSI (R = 0.323; P = .039). CONCLUSION: Patients who have undergone an ACLR continue to offload their surgical limb during both squatting and landing. Additionally, there is a relationship between movement deficits during squatting and movement deficits during landing in patients with an ACLR preparing to return to sports. CLINICAL RELEVANCE: As movement deficits during squatting and landing were related before return to sports, this study suggests that interventional approaches to improve squatting biomechanics may translate to improved landing biomechanics in patients with an ACLR.

Plantar feedback contributes to the regulation of leg stiffness.

BACKGROUND: Running and hopping involve moving in a bouncing fashion during which the limbs behave as springs. The ability to alter the stiffness of these leg springs is essential to maintaining an efficient gait. Since the plantar surface of the foot is the only part of the body to encounter the ground during bipedal locomotion, it would seem logical that some aspect of the neurological functioning of the foot is responsible for transmission of information about the surface characteristics to the central nervous system, resulting in changes in leg stiffness. METHODS: Ten subjects (9 males, 1 female) participated in this experiment. Lidocaine was injected inferior and posterior to the lateral malleolus in order to achieve tibial nerve block at the level of the ankle. Subjects hopped at 2.2 Hz on a force plate while data were collected at 1000 Hz. Data were analyzed for peak force and leg stiffness and compared using a repeated measures ANOVA. FINDINGS: Tactile sensation, deep pressure sensation, and abductor hallucis activity displayed significant decreases following the injection, as did postural stability. Subjects demonstrated a significantly decreased leg stiffness after the nerve block (P<.01). INTERPRETATION: Plantar sensation has an effect on regulating leg mechanics in hopping. A loss of sensation in this region can exert a significant impact on the properties of the leg in gait, and future research should determine the specific pathways by which plantar feedback exerts this effect.