Shallow Pockets Research Articles

Comparative Study of Allosteric GPCR Binding Sites and Their Ligandability Potential.

The steadily growing number of experimental G-protein-coupled receptor (GPCR) structures has revealed diverse locations of allosteric modulation, and yet few drugs target them. This gap highlights the need for a deeper understanding of allosteric modulation in GPCR drug discovery. The current work introduces a systematic annotation scheme to structurally classify GPCR binding sites based on receptor class, transmembrane helix contacts, and, for membrane-facing sites, membrane sublocation. This GPCR specific annotation scheme was applied to 107 GPCR structures bound by small molecules contributing to 24 distinct allosteric binding sites for comparative evaluation of three binding site detection methods (BioGPS, SiteMap, and FTMap). BioGPS identified the most in 22 of 24 sites. In addition, our property analysis showed that extrahelical allosteric ligands and binding sites represent a distinct chemical space characterized by shallow pockets with low volume, and the corresponding allosteric ligands showed an enrichment of halogens. Furthermore, we demonstrated that combining receptor and ligand similarity can be a viable method for ligandability assessment. One challenge regarding site prediction is the ligand shaping effect on the observed binding site, especially for extrahelical sites where the ligand-induced effect was most pronounced. To our knowledge, this is the first study presenting a binding site annotation scheme standardized for GPCRs, and it allows a comparison of allosteric binding sites across different receptors in an objective way. The insight from this study provides a framework for future GPCR binding site studies and highlights the potential of targeting allosteric sites for drug development.

Targeted sulfur(VI) fluoride exchange-mediated covalent modification of a tyrosine residue in the catalytic pocket of tyrosyl-DNA phosphodiesterase 1

Developing effective inhibitors of the DNA repair enzyme tyrosyl-DNA phosphodiesterase 1 (TDP1) has been challenging because of the enzyme shallow catalytic pocket and non-specific substrate binding interactions. Recently, we discovered a quinolone-binding hot spot in TDP1’s active site proximal to the evolutionary conserved Y204 and F259 residues that position DNA. Sulfur (VI) fluoride exchange (SuFEx) is a biocompatible click chemistry reaction that enables acylation of protein residues, including tyrosine. Selective protein modifications can provide insights into the biological roles of proteins and inform ligand design. As we report herein, we used SuFEx chemistries to prepare covalent TDP1-bound binders showing site-specific covalent bonds with Y204. Our work presents the first application of SuFEx chemistries to TDP1 ligands. It validates the ability to covalently modify specific TDP1 residues by designed targeting and adds to the chemical biology resource toolbox for studying TDP1.

Structural Basis for the Recognition of GPRC5D by Talquetamab, a Bispecific Antibody for Multiple Myeloma

Multiple myeloma (MM) is a complex hematological malignancy characterized by abnormal antibody production from plasma cells. Despite advances in the treatment, many patients experience disease relapse or become refractory to treatment. G-protein-coupled receptor class C group 5 member D (GPRC5D), an orphan GPCR predominantly expressed in MM cells, is emerging as a promising target for MM immunotherapy. Talquetamab, a Food and Drug Administration-approved T-cell-directing bispecific antibody developed for treatment of MM, targets GPRC5D. Here, we elucidate the structure of GPRC5D complexed with the Fab fragment of talquetamab, using cryo-electron microscopy, providing the basis for recognition of GPRC5D by the bispecific antibody. GPRC5D forms a symmetric homodimer with the interface between transmembrane helix (TM) 4 of one protomer and TM4/5 of the other protomer. A single talquetamab Fab interacts with the GPRC5D dimer with its orientation toward the dimer interface. All six complementarity-determining regions of talquetamab engage with extracellular loops and TM3/5/7. In particular, the side-chain of an arginine residue from the antibody penetrates into a shallow pocket on the extracellular surface of GPRC5D. The structure offers insights for optimizing antibody design against GPRC5D for relapsed or refractory MM therapy.

Microbiological and Salivary Biomarkers Successfully Predict Site-Specific and Whole-Mouth Outcomes of Nonsurgical Periodontal Treatment.

Background/Objectives: Nonsurgical periodontal treatment (NSPT) is the gold-standard technique for treating periodontitis. However, an individual's susceptibility or the inadequate removal of subgingival biofilms could lead to unfavorable responses to NSPT. This study aimed to assess the potential of salivary and microbiological biomarkers in predicting the site-specific and whole-mouth outcomes of NSPT. Methods: A total of 68 periodontitis patients exhibiting 1111 periodontal pockets 4 to 6 mm in depth completed the active phase of periodontal treatment. Clinical periodontal parameters, saliva, and subgingival biofilm samples were collected from each patient at baseline and three months after NSPT. A quantitative PCR assay was used to detect the presence of Fusobaterium nucleatum and Porphyromonas gingivalis in the biofilm samples. Salivary biomarkers including matrix metalloproteinase (MMP)-9, glutathione S-transferase (GST), and Annexin-1 were assayed both qualitatively (Western blot analysis) and quantitively (ELISA). Results: NSPT yielded significant improvements in all clinical parameters, including a reduction in bacterial load and decreased levels of MMP-9 together with increased concentrations of GST and Annexin-1. The binary logistic regression suggested that the overall accuracy of P. gingivalis identification, probing pocket depth, and interproximal sites was 71.1% in predicting successful site-specific outcomes. The salivary biomarker model yielded an overall accuracy of 79.4% in predicting whole-mouth outcomes following NSPT. Conclusions: At baseline, the presence of shallow periodontal pockets at interdental locations with a lower abundance of P. gingivalis is predictive of a favorable response to NSPT at the site level. Decreased salivary MMP-9 associated with increased GST and Annexin-1 levels can predict successful whole-mouth outcomes following NSPT.

Platelet-rich fibrin as an adjunct to scaling and root planing in treatment of shallow periodontal pockets: A randomized clinical trial

ObjectivesTo evaluate the efficacy of platelet-rich fibrin (PRF) as an adjunct to scaling and root planing (ScRp) for healing shallow periodontal pockets. MethodsTwelve patients with periodontitis were enrolled in this split-mouth, randomized clinical trial. A total of 24 shallow periodontal pockets (4–6 mm) were treated by either ScRp alone (control) or PRF (test). Clinical attachment loss (CAL), probing pocket depth (PPD), bleeding on probing (BOP), and plaque index (PLI), as well as platelet-derived growth factor-BB (PDGF-BB) by enzyme-linked immunosorbent assay (ELISA) in gingival crevicular fluid (GCF) were measured at baseline and at 1- and 3-month follow-up visits. ResultsAt 1- and 3-month follow-up visits, greater CAL gains (2.6 ± 0.25 mm and 3.26 ± 0.31 mm, respectively) and PPD reductions (2.58 ± 0.38 and 3.31 ± 0.39 mm, respectively) were observed in the test group compared to those in controls (CAL gain of 1.01 ± 0.49 mm and 1.43 ± 0.48 mm; PPD reduction of 1.1 ± 0.55 and 1.37 ± 0.49 mm, respectively). In addition, the increase in PDGF-BB in GCF in the test group (724.5 ± 186.09 pg/μl and 1957.5 ± 472.9 pg/μl) was significantly greater than that in controls (109.3 ± 24.07 and 614.64 ± 209.3 pg/μl) at 1- and 3-month follow-up visits, respectively. ConclusionsThe noninvasive use of PRF as an adjunct to ScRp successfully improved clinical periodontal parameters and might contribute to increased PDGF-BB in GCF.

Assessment of Oral Health Status and Treatment Needs among a Cohort of Pregnant Women in Hyderabad City – A Cross-sectional Survey

Abstract Aim: The aim of this study was to assess the oral health status and treatment needs of pregnant women visiting the Osmania Government Maternity Hospital in Hyderabad, India. Materials and Methods: A cross-sectional survey was conducted for about 5 months, encompassing 606 pregnant women. In the clinical oral examination, the community periodontal index (CPI), loss of attachment (LOA), dentition status, and treatment needs were recorded using the World Health Organization 1997 pro forma. The data were analyzed using SPSS software version 22.0. Results: On periodontal examination, 279 (46%) subjects had calculus (Code “2”), and 219 (36.1%) respondents presented with shallow pockets (Code “3”) on probing. Furthermore, more than half of the study participants (353; 58.3%) had an LOA of 4–5 mm (Code “1”). Women in the third trimester had a significantly (P < 0.001) higher mean number of sextants affected with CPI Code “1” (1.94 ± 1.08), Code “3” (1.03 ± 1.14), Code “4” (0.09 ± 0.37), and LOA Code “2” (0.13 ± 0.40). Likewise, study participants who conceived three or more times had a significantly higher mean number of sextants affected by CPI Code “3” (1.09 ± 1.14; P < 0.001), LOA Code “1” (2.22 ± 1.53; P < 0.001), and LOA Code “2” (0.12 ± 0.36; P = 0.02). The mean decayed missing filled teeth (3.91 ± 3.25, 4.00 ± 3.24) and decayed teeth (3.83 ± 3.25, 3.96 ± 3.24) showed a significant (P = 0.0001) relationship with women in the third trimester, those who conceived three or more times. Based on treatment needs, 420 (69.3%) teeth are in need of one surface filling, 217 (35.8%) teeth require two or more surface fillings, and pulp care and restoration were needed in 213 (35.1%) teeth. On a note, a greater percentage of the participants (84%) had never been to the dentist. Conclusion: The study emphasizes the need to structure preventive oral health education programs at regular intervals targeting expecting women.

Prevalence of Periodontal Disease Among Undergraduate Students In AL-Qadisiyah Governorate, Iraq: A Cross-Sectional Study

Periodontal disease, which involves gingivitis and periodontitis, is a common oral infection that affects the tissues that surround and support the teeth. Previously there were no epidemiological studies to assess the prevalence of periodontal disease in Al-Qadisiyah governorate generally and Al-Qadisiyah colleges and institute specifically. Therefore, an epidemiological study to determine the prevalence of periodontal diseases is essentially needed. To assess the prevalence of periodontal disease for the final year undergraduate students at all colleges and institute in Al-Qadisiyah governorate, Iraq and investigate the effect of age, gender, socioeconomic status, marital status and lifestyle factors on periodontal health status. A cross-sectional study was performed in AL-Qadisiyah colleges and institute. A total of 800 last stages students were examined using the community periodontal index for treatment needs and chosen randomly through stratified random sampling. A questionnaire was used to record information about socioeconomic status, marital status, and lifestyle, and correlated with periodontal health status. The overall prevalence rate of periodontal disease for the final year of undergraduate students in Al-Qadisiyah colleges and institute was 84.6%. Bleeding code 1 and calculus code 2 recorded approximately similar percentages (31.5%) and (31.4) respectively and only 15.4 % of students had healthy periodontium, while the percentage of shallow and deep pockets recorded 18.4 % and 3.4% respectively. Community periodontal index for treatment needs varied non-significantly among marital status in which single students had healthier periodontium (16.5%) compared to married. The statistical analysis also showed that students with very good socioeconomic status and good lifestyle had the highest percentages of healthy periodontium (50%), (22.6%) respectively. The present study shows a higher prevalence rate of bleeding and calculus among final year of undergraduate students in Al–Qadisiyah colleges and institute with a lack of knowledge about periodontal health status. Hence, there are significant associations of age, gender, marital status, socioeconomic status, and lifestyle with periodontal health conditions, also the results indicated that the majority of the students need preventive programs to reduce PDs initiation or progression.

The Neapolitan Yellow Tuff record in the Gaeta Gulf (Eastern Tyrrhenian margin, Southern Italy)

Abstract. The Neapolitan Yellow Tuff caldera-forming eruption (ca. 15 kyr) is a large event that occurred at Phlegrean Fields during the Late Glacial–Holocene time span. The eruptive products represent a widespread isochronous marker that links the marine and terrestrial stratigraphic record of the central Mediterranean and Northern Europe sequences. In this research, we describe the stratigraphic signature of the NYT deposits in the Late Glacial–Holocene shelf sequence of the Gulf of Gaeta (northern Campania region, southern Tyrrhenian Sea) throughout the seismo-stratigraphic interpretation of a grid of high and very high-resolution Chirp seismic profiles. The seismo-stratigraphic data allowed to individuate in the Upper Pleistocene–Holocene stratigraphic architecture a continental shelf characterized by an offlap prograding succession followed by a very thick transgressive onlapping unit, and by an upper highstand unit mainly characterized by undulations probably linked to gas-prone sediment. The NYT reflector observed was parallel and with a high amplitude. It was continuously detected from the shelf to the slope within transgressive deposits. On the inner shelf, its continuity was interrupted by shallow gas pockets. The NYT depths ranges from 2 m b.s.f. on the slope to 38 m b.s.f. on the continental shelf. The NYT event appeared to have played a key role in controlling the stratigraphic architecture of the studied area during the last transgression, acting as a trigger for the formation of the aggradational sedimentary successions fed by pyroclastic and volcaniclastic deposits.

Uncommon P1 Anchor-featured Viral T Cell Epitope Preference within HLA-A*2601 and HLA-A*0101 Individuals.

The individual HLA-related susceptibility to emerging viral diseases such as COVID-19 underscores the importance of understanding how HLA polymorphism influences peptide presentation and T cell recognition. Similar to HLA-A*0101, which is one of the earliest identified HLA alleles among the human population, HLA-A*2601 possesses a similar characteristic for the binding peptide and acts as a prevalent allomorph in HLA-I. In this study, we found that, compared with HLA-A*0101, HLA-A*2601 individuals exhibit distinctive features for the T cell responses to SARS-CoV-2 and influenza virus after infection and/or vaccination. The heterogeneous T cell responses can be attributed to the distinct preference of HLA-A*2601 and HLA-A*0101 to T cell epitope motifs with negative-charged residues at the P1 and P3 positions, respectively. Furthermore, we determined the crystal structures of the HLA-A*2601 complexed to four peptides derived from SARS-CoV-2 and human papillomavirus, with one structure of HLA-A*0101 for comparison. The shallow pocket C of HLA-A*2601 results in the promiscuous presentation of peptides with "switchable" bulged conformations because of the secondary anchor in the median portion. Notably, the hydrogen bond network formed between the negative-charged P1 anchors and the HLA-A*2601-specific residues lead to a "closed" conformation and solid placement for the P1 secondary anchor accommodation in pocket A. This insight sheds light on the intricate relationship between HLA I allelic allomorphs, peptide binding, and the immune response and provides valuable implications for understanding disease susceptibility and potential vaccine design.

Efficient Refinement of Complex Structures of Flexible Histone Peptides Using Post-Docking Molecular Dynamics Protocols.

Histones are keys to many epigenetic events and their complexes have therapeutic and diagnostic importance. The determination of the structures of histone complexes is fundamental in the design of new drugs. Computational molecular docking is widely used for the prediction of target-ligand complexes. Large, linear peptides like the tail regions of histones are challenging ligands for docking due to their large conformational flexibility, extensive hydration, and weak interactions with the shallow binding pockets of their reader proteins. Thus, fast docking methods often fail to produce complex structures of such peptide ligands at a level appropriate for drug design. To address this challenge, and improve the structural quality of the docked complexes, post-docking refinement has been applied using various molecular dynamics (MD) approaches. However, a final consensus has not been reached on the desired MD refinement protocol. In this present study, MD refinement strategies were systematically explored on a set of problematic complexes of histone peptide ligands with relatively large errors in their docked geometries. Six protocols were compared that differ in their MD simulation parameters. In all cases, pre-MD hydration of the complex interface regions was applied to avoid the unwanted presence of empty cavities. The best-performing protocol achieved a median of 32% improvement over the docked structures in terms of the change in root mean squared deviations from the experimental references. The influence of structural factors and explicit hydration on the performance of post-docking MD refinements are also discussed to help with their implementation in future methods and applications.

Efficacy of Non-Surgical Periodontal Therapy with Adjunctive Methylene Blue and Toluidine Blue O Mediated Photodynamic in Treatment of Periodontitis: A Randomized Clinical Trial.

This study aimed to examine the efficacy of methylene blue (MB) and toluidine blue O (TBO) photodynamic therapy (PDT) as adjuncts to root surface debridement (RSD). This split-mouth, randomized, controlled clinical trial included eighteen patients, and a total of 332 sites (control = 102, MB = 124 and TBO = 106) were examined. Two sessions of PDT were completed at baseline and two weeks after RSD. Clinical parameters of bleeding on probing (BOP), plaque index (PI), probing pocket depth (PPD), and clinical attachment level (CAL) were measured pre- and post-treatment. PPD and BOP reductions in sites treated by RSD with adjunctive photosensitizers (MB and TBO) were significantly higher than in control sites. RSD with MB showed higher efficacy in improving moderately deep pockets (OR 3.350), while adjunctive TBO showed better results in treating deeper pockets (OR 4.643). Results suggested that adjunctive use of MB and TBO to RSD could significantly improve periodontal pocket closure and reduce signs of inflammation. In addition, TBO seems to be more efficient in treating deep periodontal pockets than MB, which is more effective in resolving shallower pockets.

Change in clinical parameters after subgingival instrumentation for the treatment of periodontitis and timing of periodontal re-evaluation: A systematic review and meta-analysis.

To evaluate the changes in periodontal parameters (reduction in probing pocket depth [PPD], gain in clinical attachment level [CAL] and reduction in full-mouth bleeding on probing [BoP]) after subgingival instrumentation of periodontal pockets at different time points in systemically healthy patients suffering from periodontitis. Four databases were searched for RCTs that carried out subgingival instrumentation in periodontal pockets and evaluated PPD at a minimum of two consecutive time points other than baseline. The analysis was conducted for both all pocket depths and stratified for initially shallow (4-5 mm) and deep (≥6 mm) pockets and data were extracted for various time points, 1-2, 3-4 and 5-6 months. Weighted mean effects (WMEs) were calculated with 95% confidence interval (CI) and predictive intervals were calculated. Twenty-nine RCTs were identified, and all of them were included in the meta-analysis. The results showed that for both shallow and deep pockets there was a small though clinically meaningful change between 1- to 2-month and 3- to 4-month time points and between these and 5-6 months. In systemically healthy patients, the greater part of reduction in PPD and gain in CAL occurs within the first 1-2 months after subgingival instrumentation. However, additional benefits in terms of pocket depth reduction occur beyond these early time points.

Interpretasi Mekanisme Erupsi Gunung Api Lokon berdasarkan Aktivitas Kegempaan Pra-Erupsi 4 – 12 September 2014

Prior to the eruption of Lokon Volcano on September 13 2014 at 03.09 WITA, there were 62 volcanic earthquake events recorded by seismometers since September 4 2014. This research aims to interpret the eruption mechanism of Lokon Volcano based on seismic activity in the pre-eruption period. 04 - 12 September 2014. The distribution of source points for volcanic earthquakes, both hypocenters, can be visualized using the seismic method, namely by processing digital seimogram data using Swarm 3.1.0 software, GAD (Geiger's method with Adaptive Damping) and Origin 8. Data used, sourced from three stations, namely, Empung (EMP), Wailan (WLN), Kinilow (KIN). Lokon volcano. The hypocenter distribution of volcanic earthquakes is concentrated at a depth of 0 - 1,500 m above sea level and it was found that the aseismic area which is assumed to be a shallow magma pocket area from the Lokon volcano is at a depth of 1,500 - 3,000 m above sea level. The interpretation results show that the eruption mechanism of Lokon Volcano originates from disturbances in the volcanic system which are thought to be the result of tectonic earthquakes in the subduction zone. This causes the magma to rise upwards to supply shallow magma pockets until it reaches maximum pressure and an eruption occurs.

Effects of intermittent fasting on periodontal inflammation and subgingival microbiota.

Studies on the impact of intermittent fasting on periodontal health are still scarce. Thus, this study evaluated the effects of long-term intermittent fasting on periodontal health and the subgingival microbiota. This pilot study was part of a nonrandomized controlled trial. Overweight/obese participants (n=14) entered an intermittent fasting program, specifically the 5:2 diet, in which they restricted caloric intake to about a quarter of the normal total daily caloric expenditure for two nonconsecutive days/week. Subjects underwent a thorough clinical and laboratory examination, including an assessment of their periodontal condition, at baseline and 6months after starting the diet. Additionally, subgingival microbiota was assessed by 16S rRNA gene sequencing. After 6months of intermittent fasting, weight, body mass index, C-reactive protein, hemoglobin A1c (HbA1c), and the cholesterol profile improved significantly (p<0.05). Moreover, significant reductions were observed in bleeding on probing (p=0.01) and the presence of shallow periodontal pockets after fasting (p<0.001), while no significant change was seen in plaque index (p=0.14). While we did not observe significant changes in α- or β-diversity of the subgingival microbiota related to dietary intervention (p>0.05), significant differences were seen in the abundances of several taxa among individuals exhibiting ≥60% reduction (good responders) in probing pocket depth of 4-5mm compared to those with <60% reduction (bad responders). Intermittent fasting decreased systemic and periodontal inflammation. Although the subgingival microbiota was unaltered by this intervention, apparent taxonomic variability was observed between good and bad responders.

Promiscuous G-protein activation by the calcium-sensing receptor.

The human calcium-sensing receptor (CaSR) detects fluctuations in the extracellular Ca2+ concentration and maintains Ca2+ homeostasis1,2. It also mediates diverse cellular processes not associated with Ca2+ balance3-5. The functional pleiotropy of CaSR arises in part from its ability to signal through several G-protein subtypes6. We determined structures of CaSR in complex with G proteins from three different subfamilies: Gq, Gi and Gs. We found that the homodimeric CaSR of each complex couples to a single G protein through a common mode. This involves the C-terminal helix of each Gα subunit binding to a shallow pocket that is formed in one CaSR subunit by all three intracellular loops (ICL1-ICL3), an extended transmembrane helix 3 and an ordered C-terminal region. G-protein binding expands the transmembrane dimer interface, which is further stabilized by phospholipid. The restraint imposed by the receptor dimer, in combination with ICL2, enables G-protein activation by facilitating conformational transition of Gα. We identified a single Gα residue that determines Gq and Gs versus Gi selectivity. The length and flexibility of ICL2 allows CaSR to bind all three Gα subtypes, thereby conferring capacity for promiscuous G-protein coupling.

Topography prediction of high-aspect ratio features milled with overlapping abrasive slurry jet footprints considering fluid confinement effects

Overlapping adjacent passes of abrasive slurry jets can be used to fabricate micro-pockets or blind micro-slots. This study investigates the propagation of surfaces milled using overlapping passes of a high-pressure abrasive slurry jet on a ductile 6061-T6 target over a wide range of aspect ratios. Features such as shallow pockets with low aspect ratios to deep slots with aspect ratios ∼1.5 were fabricated to investigate the correlations between the aspect ratio, degree of pass overlap, and number of repeating passes with the machined surface evolution characteristics. A comprehensive model was presented that combined computational fluid dynamics (CFD) with surface evolution to predict the evolving machined feature topography after each of the overlapping and repeating passes, and to provide insights into experimental observations. The model explicitly considered the CFD-predicted local particle impact angles and velocities as well as effects due to secondary impacts, high sidewall slopes, and stagnation effects. For larger degrees of overlap, measurements revealed a more than linear rate of increase in depth after each repeat of two adjacent overlapped passes, as opposed to a linear depth increase for smaller overlaps and blind pockets. Large overlaps were found to result in asymmetric features. Up to a critical aspect ratio of ∼0.65, the model predicted the surface evolution of the features to within <8.6% of those measured. Beyond that, randomness in the machined feature shape and size made the process challenging to control and the surface evolution difficult to predict. Nevertheless, the model was able to elucidate the reasons for the randomness and other observed phenomena such as the more than linear growth in etch rate, and the occurrence of feature asymmetry. The findings emphasize that flow confinement, increases in the jet's turbulent kinetic energy, and the formation of vortices at the bottom of the machined features are critical factors influencing the machining process, and that in these cases explicit modeling approaches like the one presented in this study must be used.

Enhancing the endo-activity of the thermophilic chitinase to yield chitooligosaccharides with high degrees of polymerization

Thermophilic endo-chitinases are essential for production of highly polymerized chitooligosaccharides, which are advantageous for plant immunity, animal nutrition and health. However, thermophilic endo-chitinases are scarce and the transformation from exo- to endo-activity of chitinases is still a challenging problem. In this study, to enhance the endo-activity of the thermophilic chitinase Chi304, we proposed two approaches for rational design based on comprehensive structural and evolutionary analyses. Four effective single-point mutants were identified among 28 designed mutations. The ratio of (GlcNAc)3 to (GlcNAc)2 quantity (DP3/2) in the hydrolysates of the four single-point mutants undertaking colloidal chitin degradation were 1.89, 1.65, 1.24, and 1.38 times that of Chi304, respectively. When combining to double-point mutants, the DP3/2 proportions produced by F79A/W140R, F79A/M264L, F79A/W272R, and M264L/W272R were 2.06, 1.67, 1.82, and 1.86 times that of Chi304 and all four double-point mutants exhibited enhanced endo-activity. When applied to produce chitooligosaccharides (DP ≥ 3), F79A/W140R accumulated the most (GlcNAc)4, while M264L/W272R was the best to produce (GlcNAc)3, which was 2.28 times that of Chi304. The two mutants had exposed shallower substrate-binding pockets and stronger binding abilities to shape the substrate. Overall, this research offers a practical approach to altering the cutting pattern of a chitinase to generate functional chitooligosaccharides.Graphical abstract

A mixed method study to assess orodental problems and barriers to utilisation of orodental care in pregnant ladies of Himachal Pradesh

Background: The hormonal changes along with decreased salivation during pregnancy result in many orodental problems. The existing orodental conditions worsen due to barriers to the utilisation of orodental care during pregnancy. The objective of this study is to assess orodental problems and barriers to utilisation of orodenatl care in pregnant ladies Methods: Sample size is calculated using Cochran’s formula. The Purposive sampling technique was used to select hospitals and pregnant women from OPD of the gynecology and obstetrics department of hospitals selected in Himachal Pradesh. Using DMFT and CPI index, interview schedules the study's specific objectives are achieved. Results: Out of a sample of 112, 92.8% of pregnant women had dental caries, 34.8% of pregnant women had got restorative treatment and 25.8% had their teeth extracted. The majority of pregnant women (51.8%) had dental calculus and (29.5%) shallow periodontal pockets. The system, personal, and caregiver-related barriers are responsible for the underutilisation of orodental care among pregnant women. Conclusions: This study concluded many pregnant women face orodental problems along with a major factor of lack of dental education and no dental insurance associated with their orodental problems.

Limited high-throughput screening compatibility of the phenuivirus cap-binding domain

Bunyaviruses constitute a large and diverse group of viruses encompassing many emerging pathogens, such as Rift Valley fever virus (family Phenuiviridae), with public and veterinary health relevance but with very limited medical countermeasures are available. For the development of antiviral strategies, the identification and validation of virus-specific targets would be of high value. The cap-snatching mechanism is an essential process in the life cycle of bunyaviruses to produce capped mRNAs, which are then recognized and translated into viral proteins by the host cell translation machinery. Cap-snatching involves cap-binding as well as endonuclease functions and both activities have been demonstrated to be druggable in related influenza viruses. Here, we explore the suitability of the phenuivirus cap-binding function as a target in medium- and high-throughput drug discovery approaches. We developed a range of in vitro assays aiming to detect the interaction between the cap-binding domain (CBD) and the analogue of its natural cap-ligand m7GTP. However, constricted by its shallow binding pocket and low affinity for m7GTP, we conclude that the CBD has limited small molecule targeting potential using classical in vitro drug discovery approaches.

Periodontal Care In Prolanis Diabetics: Sungai Ulin and South Banjarbaru Primary Health Care Study

The research focused on oral health among diabetes mellitus patients, particularly in preventing periodontal disease within the prolanis group. Using analytical descriptive methods, the study evaluated oral cavity abnormalities. Findings at Sungai Ulin and South Banjarbaru Community Health Centers revealed a significant number of respondents with tartar and shallow gum pockets, indicating a substantial need for care. The majority scored 2 on the CPITN scale (61.8%), underscoring the demand for attention from healthcare providers including community health workers, dentists, and dental nurses. Recommendations stress the pivotal role of oral health maintenance for diabetes mellitus individuals. These suggestions encompass regular blood sugar monitoring, consistent dental hygiene, tartar removal, denture care, and eliminating local factors causing gum infections. Periodontal disease's impact on daily life includes chewing difficulties, speech impediments, and tooth loss, accentuating the urgency of preventive measures for diabetes mellitus patients. In summary, proactive preventive actions are crucial to address periodontal disease among diabetes mellitus individuals, aiming to diminish its occurrence and improve their overall well-being and quality of life.