Identifying SARS-CoV-2 antiviral compounds by screening for small molecule inhibitors of Nsp5 main protease.

The coronavirus 2019 (COVID-19) pandemic, caused by the severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2), spread around the world with unprecedented health and socio-economic effects for the global population. While different vaccines are now being made available, very few antiviral drugs have been approved. The main viral protease (nsp5) of SARS-CoV-2 provides an excellent target for antivirals, due to its essential and conserved function in the viral replication cycle. We have expressed, purified and developed assays for nsp5 protease activity. We screened the nsp5 protease against a custom chemical library of over 5000 characterised pharmaceuticals. We identified calpain inhibitor I and three different peptidyl fluoromethylketones (FMK) as inhibitors of nsp5 activity in vitro, with IC50 values in the low micromolar range. By altering the sequence of our peptidomimetic FMK inhibitors to better mimic the substrate sequence of nsp5, we generated an inhibitor with a subnanomolar IC50. Calpain inhibitor I inhibited viral infection in monkey-derived Vero E6 cells, with an EC50 in the low micromolar range. The most potent and commercially available peptidyl-FMK compound inhibited viral growth in Vero E6 cells to some extent, while our custom peptidyl FMK inhibitor offered a marked antiviral improvement.

Assessment of dementia knowledge in Indian speech-language pathology students.

OBJECTIVES: Speech-language pathologists (SLPs) have a crucial role in assisting individuals with dementia due to the communication and swallowing challenges associated with the disease. As the number of dementia cases rises in India at an increasing rate, investigating the level of dementia knowledge of SLP students can offer insight into the preparedness of the healthcare system to meet this emerging demand. METHOD: A cross-sectional survey was conducted on SLP students pursuing their final year undergraduate, postgraduate and doctoral degrees from four universities across India. Dementia knowledge was assessed using the Dementia Knowledge Assessment Scale (DKAS) and information about previous dementia exposure (both formal and informal) was collected. The collected data were analysed using quantitative methods. RESULTS: A total of 220 students (64.70% response rate) completed the survey. Overall dementia knowledge was inadequate with an average score of 22.08 ± 10.06. Previous dementia exposure among the students was also found to be low and did not affect dementia knowledge scores. DISCUSSION: Despite the fundamental role SLPs play in the care of individuals with dementia, the lack of knowledge in this area emphasizes the need for enhancing dementia training programs through educational curricula and clinical placements.

Peptide Therapeutics and the Pharmaceutical Industry: Barriers Encountered Translating from the Laboratory to Patients.

Peptides are receiving increasing interest as clinical therapeutics. These highly tunable molecules can be tailored to achieve desirable biocompatibility and biodegradability with simultaneously selective and potent therapeutic effects. Despite challenges regarding up-scaling and licensing of peptide products, their vast clinical potential is reflected in the 60 plus peptide-based therapeutics already on the market, and the further 500 derivatives currently in developmental stages. Peptides are proving effective for a multitude of disease states including: type 2 diabetes (controlled using the licensed glucagon-like peptide-1 receptor liraglutide); irritable bowel syndrome managed with linaclotide (currently at approval stages); acromegaly (treated with octapeptide somatostatin analogues lanreotide and octreotide); selective or broad spectrum microbicidal agents such as the Gram-positive selective PTP-7 and antifungal heliomicin; anticancer agents including goserelin used as either adjuvant or monotherapy for prostate and breast cancer, and the first marketed peptide derived vaccine against prostate cancer, sipuleucel-T. Research is also focusing on improving the biostability of peptides. This is achieved through a number of mechanisms ranging from replacement of naturally occurring L-amino acid enantiomers with D-amino acid forms, lipidation, peptidomimetics, N-methylation, cyclization and exploitation of carrier systems. The development of self-assembling peptides are paving the way for sustained release peptide formulations and already two such licensed examples exist, lanreotide and octreotide. The versatility and tunability of peptide-based products is resulting in increased translation of peptide therapies, however significant challenges remain with regard to their wider implementation. This review highlights some of the notable peptide therapeutics discovered to date and the difficulties encountered by the pharmaceutical industry in translating these molecules to the clinical setting for patient benefit, providing some possible solutions to the most challenging barriers.

Evaluation of Shear Bond Strength of Newer Bonding Systems on Superficial and Deep Dentin.

BACKGROUND: The purpose of this study was to compare the shear bond strength of nanocomposite resin to superficial dentin and deep dentin using two different dentin bonding systems. MATERIALS AND METHODS: All teeth were sectioned at various levels (superficial dentin: Dentin within 0.5-1 mm of dentinoenamel junction; deep dentin: Dentin within 0.5 mm of the highest pulp horn) using a Carborundum Disc and embedded in acrylic block of specific size. Selected specimens (60 premolar teeth) were grouped randomly into three groups, the groups were differentiated into superficial dentin, deep dentin, and control group which were further divided into sub Group A and Subgroup B containing 10 teeth each, depending on the bonding agents used. In Subgroup A, Tetric N Bond, and in Subgroup B Single Bond Universal were used. In the control group no bonding agent was used. The specimens were thermocycled for 500 cycles between 5°C and 55°C water bath for 40 s. Finally, the specimens were subjected to shear bond strength study under INSTRON machine (Universal Testing Machine). The maximum shear bond strengths were noted at the time of fracture (de-bonding) of the restorative material. Results were analyzed using ANOVA test, Bonferroni test, and paired t-test. RESULTS: Bond strength values of fifth generation bonding system (Tetric N Bond) showed higher mean shear bond strength compared to seventh generation bonding system (Single Bond Universal). There was a significant fall in bond strength values as one reaches deeper levels of dentin from superficial to deep dentin. CONCLUSION: There was a significant difference between the bond strength of fifth generation bonding system (Tetric N Bond) and seventh generation bonding system (Single Bond Universal). Decrease in the bond strength values is seen for the deeper level of dentin as compared to superficial dentin.

Evaluation of the Effect of Surface Polishing, Oral Beverages and Food Colorants on Color Stability and Surface Roughness of Nanocomposite Resins.

BACKGROUND: It is beyond doubt that finishing and polishing of a composite restoration enhance its esthetics and, is also essential for the health of the periodontium. A variety of instruments are commonly used for finishing and polishing tooth-colored restorative materials Thus, it is important to understand which type of surface finishing treatments would significantly affect the staining and surface irregularities of the composite resin restoration. Still one of the properties of the composite resins that have to pass the test of time is its color stability. In modern day dentistry, a large emphasis is laid over esthetics. Hence, it is important to understand the various agents capable of adversely affecting the esthetics of a restoration due to its staining capacity. Thus, the aim of this in vitro study was to evaluate the effect of surface polishing, oral beverages and food colorants on the color stability and surface roughness of nanocomposite resins. MATERIALS AND METHODS: 90 Disks of nanocomposites resin (Filtek Z350 XT) measuring 8 mm in diameter and 2 mm in thickness were fabricated using a custom made silicon mold. Pre-polishing surface roughness (Ra1) of all the 90 samples were measured using a Surface Profilometer. The nano-composite disks were then randomly divided into 3 groups with 30 samples in each group. Group I: CONTROL GROUP: The samples were not subjected to any polishing procedures. Group II: Sof-Lex group: Samples subjected to polishing using different grits of Sof-Lex disks. Group III: Diamond polishing paste group: Samples were subjected with a polishing paste consisting of diamond particles. Following polishing procedures, the surface roughness of all samples were measured again to obtain change in surface roughness due to polishing procedures (Ra2), pre immersion spectrophotometric value (ΔE1) was also recorded for baseline color of the samples. The samples were then divided into subgroups (A, B, C, D, E), by including every first sample in Subgroup A, second in Subgroup B, third in Subgroup C, fourth in Subgroup D, and fifth in Subgroup E. Each was immersed in the respective test solution for 10 min, twice a day for 30 days. Group A - Tea, Group B - Coffee, Group C - Cola, Group D - Turmeric, Group E - Control (artificial saliva). Post immersion profilometric value was recorded to evaluate roughness bought about by the solutions (Ra3) and spectrophotometric value was recorded to evaluate the color change in samples (ΔE2). Results were statistically analyzed using ANOVA. RESULTS: Higher mean roughness (Ra2-Ra1) value was recorded in Sof-Lex, followed by Diamond polishing paste and Control group. Comparison of surface roughness caused due to beverages and food colorant solution showed subgroup C (Coca Cola) increased surface roughness in all groups (Group I, II, III). Subgroup D (Turmeric) had the highest discoloration potential (P < 0001) in all groups, followed by coffee, tea, coca-cola and artificial saliva. Sof-Lex polishing System showed most color stability. CONCLUSION: Polishing procedures significantly roughen the surface of the restoration compared to the unpolished Mylar controls. One-step polishing system (diamond polishing paste) produces a smoother surface compared to a multi-step system (Sof-Lex polishing disks). Turmeric solution caused maximum staining of the samples, to a visually perceptible level.

Comparative Evaluation of Bond Strength of Dual-Cured Resin Cements: An In-Vitro Study.

BACKGROUND: To compare the microtensile bond strength of resin cements to enamel and dentin and to determine the type of bond failure using stereomicroscope. MATERIALS AND METHODS: In this in-vitro study 40 teeth were embedded in acrylic resin and divided into two main groups i.e., Group A for enamel and Group B for dentin. Each group is again subdivided into four subgroups, which are as follows; Subgroup 1 for Calibra resin cement, Subgroup 2 for Paracem, Subgroup 3 for Variolink II and Subgroup 4 for Rely X ARC. These resin cements were applied on enamel and dentin according to manufacturer's instructions followed by incremental build-up of composite resin on the top of resin cements. Each tooth was sectioned perpendicular to the resin-substrate interface with a slow speed diamond saw under water cooling yielding sections of approximately 1 mm(2). On an average, three sections from each tooth were used for testing. The beams obtained after sectioning were stressed to failure under tension in a custom made stainless steel forceps held in a universal testing machine (Lloyd) at a crosshead speed of 1.0 mm/min. Results were analyzed using two-way analysis of variance, independent t-test, and Tukey's HSD post-hoc test. RESULTS: Cements bonded to enamel substrates showed higher mean bond strength compared to dentin, which is statistically significant. Rely X ARC showed highest mean bond strength to both the substrates. CONCLUSION: There was a significant difference between the bond strength to enamel and dentin and, Rely X ARC resin cement showed higher bond strength compared with the other groups.