Spatially modulated erbium YAG laser as a treatment for diabetic ulcer.

OBJECTIVE: Diabetic ulcers are a significant healthcare challenge, capable of diminishing quality of life, lengthening hospitalisation stay, and incurring substantial costs for patients and healthcare systems. Erbium-doped yttrium-aluminum-garnet (Er-YAG) laser has been evolving as a prospective intervention for addressing wounds of various aetiologies. Despite this, the literature remains limited in appraising the effectiveness of laser therapy specifically in diabetic wounds. This study investigates the impact of employing a spatially modulated Er-YAG laser as a therapeutic approach for treating diabetic ulcers. METHOD: In a single-arm study conducted from November 2017 to April 2023, patients with hard-to-heal ulcers were treated in a two-step approach of wound debridement using Er-YAG laser for ablation and biostimulation through deep tissue resonance using RecoSMA (Multiline laser system, LINLINE MS, Latvia) laser technology. Ulcers received weekly laser treatment, together with routine care until healing occurred and were then followed up to observe any recurrence. The primary outcome measure was wound closure; the secondary outcome measures were time to closure, and the number of laser treatments required. Data related to sociodemographic details, size and number of diabetic ulcers, and number of sessions related to laser treatment were collected using a predesigned, pretested questionnaire before initiating the treatment. RESULTS: A total of 59 patients attending the clinic during the study period with diabetic ulcers were included in the study. The mean wound surface area at baseline was 25.7cm2 (median: 12cm2). The average number of sessions of laser treatment required was 4.41, ranging from 1-11. The size of the ulcer reduced with each session of laser treatment. The diabetic ulcers healed completely at the end of the treatment, indicating the effectiveness of the Er-YAG/RecoSMA two-step approach. CONCLUSION: Spatially modulated erbium YAG laser is effective as a therapeutic approach for treating diabetic ulcers.

Potential therapeutic effects of naringin loaded PLGA nanoparticles for the management of Alzheimer's disease: In vitro, ex vivo and in vivo investigation.

Background: Alzheimer's disease (AD) is the most prevalent type of dementia which has been affected to more the 44 million people globally. It is distinguished by gradually deteriorating memory and other cognitive abilities that precede dementia. Present treatment of AD mainly focuses on symptomatic slowing the evolution of the disease which is associated with numerous side effects such as dizziness, tiredness, nausea, vomiting, heart attack, and stroke etc. Henceforth; there is urgent need to identify the alternative treatment for management of AD. Herbal medicines have been used from long time to treat AD. One of such leading Phyto molecule is Naringin. It showed promising results against AD but suffers from poor bioavailability and require in high dose to cross the blood brain barrier. Objectives: The main objectives of proposed work are to increase the bioavailability of naringin in brain by developing Nano-suspension and preclinical evaluation of neuroprotective effect of Naringin Nano-suspension (NNS) against Scopolamine induced Alzheimer's disease in rats. Methods: The present study deals with the development, characterization of NNSand to evaluate neuroprotective effect of NNS. Nanoparticles of drug were formed by using PLGA polymer and optimized by using 32 factorial design. Optimized batch was further characterized by scanning electron microscopy (SEM) and X-ray diffraction (XRD). Further the effectiveness of NNS was preclinically investigated by performing AOTstudy as per OECD guideline 420. AD induced Albino Wistar Rats were treated with NNS orally for 14 days and then evaluated for parameters like Gross examination of brain, Relative brain weight determination, behavioural parameters, neuro-inflammatory parameters and immune-histology. Results: Optimization was carried out to study the effect of polymer concentration and number of HPH cycles on Particle size, Poly dispersity index (PDI) and % entrapment efficiency. Desirability search approach was used to select the optimized formulation. Based on the selection criteria, batch F6 having 357.6 ± 05 nm particle size, 0.168 ± 0.04 PDI and 91 ± 2% EE was selected as optimized batch. SEM analysis showed spherical morphology and XRD confirmed the molecular dispersion. Pre-treatment with NNS showed neuroprotective activity basedon results of behavioural studies, biochemical estimation, neuroinflammatory parameters and immunohistochemistry evaluations. Conclusion: As NNS showed significant neuroprotective and anti-neuro-inflammatory effect, this study opens up new ways to exploit Naringin for various therapeutic and restorative purposes.

Are there double knots in proteins? Prediction and in vitro verification based on TrmD-Tm1570 fusion from C. nitroreducens.

We have been aware of the existence of knotted proteins for over 30 years-but it is hard to predict what is the most complicated knot that can be formed in proteins. Here, we show new and the most complex knotted topologies recorded to date-double trefoil knots (31 #31). We found five domain arrangements (architectures) that result in a doubly knotted structure in almost a thousand proteins. The double knot topology is found in knotted membrane proteins from the CaCA family, that function as ion transporters, in the group of carbonic anhydrases that catalyze the hydration of carbon dioxide, and in the proteins from the SPOUT superfamily that gathers 31 knotted methyltransferases with the active site-forming knot. For each family, we predict the presence of a double knot using AlphaFold and RoseTTaFold structure prediction. In the case of the TrmD-Tm1570 protein, which is a member of SPOUT superfamily, we show that it folds in vitro and is biologically active. Our results show that this protein forms a homodimeric structure and retains the ability to modify tRNA, which is the function of the single-domain TrmD protein. However, how the protein folds and is degraded remains unknown.

Nose to brain delivery of nanosuspensions with first line antiviral agents is alternative treatment option to Neuro-AIDS treatment.

Intranasal drug delivery is one of the uprising areas of the research in targeting drug to the brain. Nose to brain drug delivery follows the olfactory pathway and purportedly known to be more efficient to deliver neuro-therapeutics to the brain by circumventing the BBB and thereby increasing bioavailability of drugs in the brain. The advantage of this method is non-invasiveness, rapid onset of action and helps to achieve site specific delivery. In this research work nanosuspension were prepared using combination of antiretroviral agents for Neuro-AIDS treatment. Nanosuspensions were prepared by high-speed homogenization, wet milling and high-pressure homogenization techniques. Formulations were analysed by SEM, FTIR, and DSC. Morphology and stability analysis was done by analysing zeta potential, particle size, and PDI. Ex-vivo diffusion study and histopathological analysis was performed using goat nasal mucosa. High pressure homogenization was found to be best technique for formulation of nanosuspension. Antiviral drugs could be delivered successfully by optimizing nasal dosage form.

Mannich Bases: Centrality in Cytotoxic Drug Design.

Mannich bases identified by Professor Carl Mannich have been the most extensively explored scaffolds for more than 100 years now. The versatile biological roles that they play have promoted their applications in many clinical conditions. The present review highlights the application of Mannich bases as cytotoxic agents, categorizing them into synthetic, semisynthetic, and prodrugs classes, and gives an exhaustive account of the work reported in the last two decades. The methods of synthesis of these cytotoxic agents, their anti-cancer potential in various cell lines, and promising leads for future drug development have also been discussed. Structure-activity relationships, along with the targets on which these cytotoxic Mannich bases act, have been included as well.

Residue conservation elucidates the evolution of r-proteins in ribosomal assembly and function.

Ribosomes are the translational machineries having two unequal subunits, small subunit (SSU) and large subunit (LSU) across all the domains of life. Origin and evolution of ribosome are encoded in its structure, and the core of the ribosome is highly conserved. Here, we have used Shannon entropy to analyze the evolution of ribosomal proteins (r-proteins) across the three domains of life. Moreover, we have analyzed the residue conservation at protein-protein (PP) and protein-RNA (PR) interfaces in SSU and LSU. Furthermore, we have studied the evolution of early, intermediate and late binding r-proteins. We show that the r-proteins of Thermus thermophilus are better conserved during the evolution. Furthermore, we find the late binders are better conserved than the early and the intermediate binders. The residues at the interior of the r-proteins are the most conserved followed by those at the interface and the solvent accessible surface. Additionally, we show that the residues at the PP interfaces are better conserved than those at the PR interfaces. However, between PR and PP interfaces, the multi-interface residues at the former are better conserved than those at the latter ones. Our findings may provide insights into the evolution of r-proteins in ribosomal assembly and function.

Dissecting macromolecular recognition sites in ribosome: implication to its self-assembly.

Interactions between macromolecules play a crucial role in ribosome assembly that follows a highly coordinated process involving RNA folding and binding of ribosomal proteins (r-proteins). Although extensive studies have been carried out to understand macromolecular interactions in ribosomes, most of them are confined to either large or small ribosomal-subunit of few species. A comparative analysis of macromolecular interactions across different domains is still missing. We have analyzed the structural and physicochemical properties of protein-protein (PP), protein-RNA (PR) and RNA-RNA (RR) interfaces in small and large subunits of ribosomes, as well as in between the two subunits. Additionally, we have also developed Random Forest (RF) classifier to catalog the r-proteins. We find significant differences as well as similarities in macromolecular recognition sites between ribosomal assemblies of prokaryotes and eukaryotes. PR interfaces are substantially larger and have more ionic interactions than PP and RR interfaces in both prokaryotes and eukaryotes. PP, PR and RR interfaces in eukaryotes are well packed compared to those in prokaryotes. However, the packing density between the large and the small subunit interfaces in the entire assembly is strikingly low in both prokaryotes and eukaryotes, indicating the periodic association and dissociation of the two subunits during the translation. The structural and physicochemical properties of PR interfaces are used to predict the r-proteins in the assembly pathway into early, intermediate and late binders using RF classifier with an accuracy of 80%. The results provide new insights into the classification of r-proteins in the assembly pathway.

Dissecting protein-protein interactions in proteasome assembly: Implication to its self-assembly.

The 26S proteasome is a multi-catalytic ATP-dependent protease complex that recognizes and cleaves damaged or misfolded proteins to maintain cellular homeostasis. The 26S subunit consists of 20S core and 19S regulatory particles. 20S core particle consists of a stack of heptameric alpha and beta subunits. To elucidate the structure-function relationship, we have dissected protein-protein interfaces of 20S core particle and analyzed structural and physiochemical properties of intra-alpha, intra-beta, inter-beta, and alpha-beta interfaces. Furthermore, we have studied the evolutionary conservation of 20S core particle. We find the size of intra-alpha interfaces is significantly larger and is more hydrophobic compared with other interfaces. Inter-beta interfaces are well packed, more polar, and have higher salt-bridge density than other interfaces. In proteasome assembly, residues in beta subunits are better conserved than alpha subunits, while multi-interface residues are the most conserved. Among all the residues at the interfaces of both alpha and beta subunits, Gly is highly conserved. The largest size of intra-alpha interfaces complies with the hypothesis that large interfaces form first during the 20S assembly. The tight packing of inter-beta interfaces makes the core particle impenetrable from outer wall of the cylinder. Comparing the three domains, eukaryotes have large and well-packed interfaces followed by archaea and bacteria. Our findings provide a structural basis of assembly of 20S core particle in all the three domains of life.

Modified Activation Technique for Nasal Stent of Nasoalveolar Molding Appliance for Columellar Lengthening in Bilateral Cleft Lip/Palate.

Bilateral cleft lip/cleft palate is associated with nasal deformities typified by a short columella. The presurgical nasoalveolar molding (NAM) therapy approach includes reduction of the size of the intraoral alveolar cleft as well as positioning of the surrounding deformed soft tissues and cartilages. In a bilateral cleft patient, NAM, along with columellar elongation, eliminates the need for columellar lengthening surgery. Thus the frequent surgical intervention to achieve the desired esthetic results can be avoided. This article proposes a modified activation technique of the nasal stent for a NAM appliance for columellar lengthening in bilateral cleft lip/palate patients. The design highlights relining of the columellar portion of the nasal stent and the wire-bending of the nasal stent to achieve desirable results within the limited span of plasticity of the nasal cartilages. With this technique the vertical taping of the premaxilla to the oral plate can be avoided.

Simple Technique to Reorient Different Casts of Same Patient on a Dental Surveyor.

Accurate planning for the framework design of removable partial dentures requires careful analysis of the diagnostic cast with a dental surveyor to determine the optimal path of placement. Some techniques described in the literature are helpful in reorienting the same cast on the surveyor, including the tripod marking method; however, there is a possibility of introducing human errors during marking and repositioning of the tripod points on to the different casts at the same location. Other techniques, which do not require markings on the cast to reorient different casts of the same patient, need specific devices or trays. This article suggests the direct use of a putty-elastomeric orientation index that can be preserved and used multiple times while reorienting different casts of the same patients at various laboratory steps. A putty elastomeric impression material is mixed and adapted on to the diagnostic cast, covering key teeth areas of the cast and incorporating the analyzing rod of the surveyor. Thus there is no need to use a special device or the tray to reorient different casts.

Host Cell Plasma Membrane Phosphatidylserine Regulates the Assembly and Budding of Ebola Virus.

UNLABELLED: Lipid-enveloped viruses replicate and bud from the host cell where they acquire their lipid coat. Ebola virus, which buds from the plasma membrane of the host cell, causes viral hemorrhagic fever and has a high fatality rate. To date, little has been known about how budding and egress of Ebola virus are mediated at the plasma membrane. We have found that the lipid phosphatidylserine (PS) regulates the assembly of Ebola virus matrix protein VP40. VP40 binds PS-containing membranes with nanomolar affinity, and binding of PS regulates VP40 localization and oligomerization on the plasma membrane inner leaflet. Further, alteration of PS levels in mammalian cells inhibits assembly and egress of VP40. Notably, interactions of VP40 with the plasma membrane induced exposure of PS on the outer leaflet of the plasma membrane at sites of egress, whereas PS is typically found only on the inner leaflet. Taking the data together, we present a model accounting for the role of plasma membrane PS in assembly of Ebola virus-like particles. IMPORTANCE: The lipid-enveloped Ebola virus causes severe infection with a high mortality rate and currently lacks FDA-approved therapeutics or vaccines. Ebola virus harbors just seven genes in its genome, and there is a critical requirement for acquisition of its lipid envelope from the plasma membrane of the human cell that it infects during the replication process. There is, however, a dearth of information available on the required contents of this envelope for egress and subsequent attachment and entry. Here we demonstrate that plasma membrane phosphatidylserine is critical for Ebola virus budding from the host cell plasma membrane. This report, to our knowledge, is the first to highlight the role of lipids in human cell membranes in the Ebola virus replication cycle and draws a clear link between selective binding and transport of a lipid across the membrane of the human cell and use of that lipid for subsequent viral entry.

Molecular architecture of protein-RNA recognition sites.

The molecular architecture of protein-RNA interfaces are analyzed using a non-redundant dataset of 152 protein-RNA complexes. We find that an average protein-RNA interface is smaller than an average protein-DNA interface but larger than an average protein-protein interface. Among the different classes of protein-RNA complexes, interfaces with tRNA are the largest, while the interfaces with the single-stranded RNA are the smallest. Significantly, RNA contributes more to the interface area than its partner protein. Moreover, unlike protein-protein interfaces where the side chain contributes less to the interface area compared to the main chain, the main chain and side chain contributions flipped in protein-RNA interfaces. We find that the protein surface in contact with the RNA in protein-RNA complexes is better packed than that in contact with the DNA in protein-DNA complexes, but loosely packed than that in contact with the protein in protein-protein complexes. Shape complementarity and electrostatic potential are the two major factors that determine the specificity of the protein-RNA interaction. We find that the H-bond density at the protein-RNA interfaces is similar with that of protein-DNA interfaces but higher than the protein-protein interfaces. Unlike protein-DNA interfaces where the deoxyribose has little role in intermolecular H-bonds, due to the presence of an oxygen atom at the 2' position, the ribose in RNA plays significant role in protein-RNA H-bonds. We find that besides H-bonds, salt bridges and stacking interactions also play significant role in stabilizing protein-nucleic acids interfaces; however, their contribution at the protein-protein interfaces is insignificant.

The Ebola virus matrix protein VP40 selectively induces vesiculation from phosphatidylserine-enriched membranes.

Ebola virus is from the Filoviridae family of viruses and is one of the most virulent pathogens known with ∼ 60% clinical fatality. The Ebola virus negative sense RNA genome encodes seven proteins including viral matrix protein 40 (VP40), which is the most abundant protein found in the virions. Within infected cells VP40 localizes at the inner leaflet of the plasma membrane (PM), binds lipids, and regulates formation of new virus particles. Expression of VP40 in mammalian cells is sufficient to form virus-like particles that are nearly indistinguishable from the authentic virions. However, how VP40 interacts with the PM and forms virus-like particles is for the most part unknown. To investigate VP40 lipid specificity in a model of viral egress we employed giant unilamellar vesicles with different lipid compositions. The results demonstrate VP40 selectively induces vesiculation from membranes containing phosphatidylserine (PS) at concentrations of PS that are representative of the PM inner leaflet content. The formation of intraluminal vesicles was not significantly detected in the presence of other important PM lipids including cholesterol and polyvalent phosphoinositides, further demonstrating PS selectivity. Taken together, these studies suggest that PM phosphatidylserine may be an important component of Ebola virus budding and that VP40 may be able to mediate PM scission.

A loop region in the N-terminal domain of Ebola virus VP40 is important in viral assembly, budding, and egress.

Ebola virus (EBOV) causes viral hemorrhagic fever in humans and can have clinical fatality rates of ~60%. The EBOV genome consists of negative sense RNA that encodes seven proteins including viral protein 40 (VP40). VP40 is the major Ebola virus matrix protein and regulates assembly and egress of infectious Ebola virus particles. It is well established that VP40 assembles on the inner leaflet of the plasma membrane of human cells to regulate viral budding where VP40 can produce virus like particles (VLPs) without other Ebola virus proteins present. The mechanistic details, however, of VP40 lipid-interactions and protein-protein interactions that are important for viral release remain to be elucidated. Here, we mutated a loop region in the N-terminal domain of VP40 (Lys127, Thr129, and Asn130) and find that mutations (K127A, T129A, and N130A) in this loop region reduce plasma membrane localization of VP40. Additionally, using total internal reflection fluorescence microscopy and number and brightness analysis we demonstrate these mutations greatly reduce VP40 oligomerization. Lastly, VLP assays demonstrate these mutations significantly reduce VLP release from cells. Taken together, these studies identify an important loop region in VP40 that may be essential to viral egress.

Effectiveness of different tooth brushing techniques on the removal of dental plaque in 6-8 year old children of Gulbarga.

OBJECTIVE: This study was undertaken to evaluate the respective effectiveness of the horizontal scrub, Fones, and modified Bass methods demonstrated on the cast to individual child within the classroom setting. MATERIALS AND METHODS: A total of 180 healthy children studying in 1(st) and 2(nd) grades in the age range of 6-8 years were randomly selected from various schools of Gulbarga district, Karnataka, India. They were equally divided into three groups. Children in each group were demonstrated only one of the three brushing techniques, viz. horizontal scrub technique to group A, Fones technique to group B, and modified Bass technique to group C, using a cast model. All the children were re-examined and reviewed after 24 h and plaque index was re-assessed to obtain the follow-up data. The results were compared with the baseline data, and statistical analysis was carried out using paired t-test and intergroup comparison was made using analysis of variance (ANOVA) test. RESULTS: Statistically significant (P < 0.001) reduction in plaque score was seen in modified Bass technique followed by horizontal scrub technique and the least efficacy was seen in Fones technique. CONCLUSION: This study showed that modified Bass technique was the most effective brushing technique in children.

Multidisciplinary treatment approach to restore deep horizontally fractured maxillary central incisor.

This case report demonstrates sequential periodontic, orthodontic and prosthodontic treatment modalities to save and restore deep horizontally fractured maxillary central incisor. The location of fracture was deep in the mucosa which reveals less than 2 mm of tooth structure to receive the crown. The procedures like surgical crown lengthening, endodontic post placement, orthodontic forced eruption, core build-up and metal-ceramic crown restoration were sequentially performed to conserve the fractured tooth. Forced eruption is preferred to surgical removal of supporting alveolar bone, since forced eruption preserves the biologic width, maintains esthetics, and at the same time exposes sound tooth structure for the placement of restorative margins.

Amelogenesis imperfecta with multiple impacted teeth and skeletal class III malocclusion: complete mouth rehabilitation of a young adult.

Amelogenesis imperfecta is an autosomal dominant disorder. It is a group of hereditary diseases showing abnormal enamel density and crown malformation. This clinical report describes the oral rehabilitation of a young adult diagnosed with a variant of hypoplastic amelogenesis imperfecta with multiple impacted teeth and skeletal class III malocclusion. The treatment procedures of teeth extractions, endodontic treatment of remaining teeth followed by post and core restorations, esthetic and functional crown lengthening, and metal ceramic fixed dental prostheses were performed sequentially in the maxillary arch. The mandibular arch was restored with an overdenture. One-year follow-up revealed satisfactory results.

Effect of Varying Layers of Two Die Spacers on Precementation Space of Full Coverage Restorations.

The purpose of this study was to evaluate and compare the effect of varying layers of two commercially available die spacers on pre-cementation space of full coverage restorations in vitro and in vivo. Seven dies were prepared for each of 15 subjects. On three dies 1, 2, 3 layers of Pico-fit and on other three dies 1, 2, 3 layers of Yeti die spacers applied, wax pattern fabricated, invested and cast. Metal copings seated in vitro on die without die spacer and on prepared tooth of respective subject with fit-checker. Thickness of fit checker was measured using micrometer at mid-axial, mid-occlusal and near finish line locations that provided pre-cementation space. Result of ANOVA tests suggested significant difference among groups with varying layers. There was no significant difference between pre-cementation space achieved with Pico-fit and Yeti die spacers. The r values suggested positive correlation between the respective pair of in vivo and in vitro groups. (1) There was significant difference between pre-cementation space at mid-axial and mid-occlusal sites achieved with 1, 2 and 3 layers of die spacers except between 1 and 2 layers and 1 and 3 layers at mid-occlusal site. (2) Pre-cementation space achieved with Pico-fit and Yeti die spacers did not differ significantly for same location, layers and in vitro and in vivo. (3) Pre-cementation space achieved in vitro was analogous to pre-cementation space achieved in vivo for respective location, layers and die spacer.

Comparative study of shade matching performance of dental students under natural daylight and daylight lamp conditions.

BACKGROUND: As visual shade matching is subject to light source variables, this study was conducted to compare shade matching performance of dental students under two lighting conditions, ie, natural daylight and a commercially available daylight lamp. MATERIALS AND METHODS: Two sets of porcelain discs were prepared. The first set consisted of eight porcelain discs of shades A2, A3, A3.5, B2, B3, C2, C3, and D3 of the Vitapan Classical Shade Guide system. The second set consisted of three porcelain discs of shade A2, B2, and C2, having exactly similar L*a*b* values to those of the corresponding shade discs in the first set. Forty dental students were asked to find the closest match for the shades A2, B2, and C2 in the second set from the first set under natural daylight and daylight lamp conditions. The average ΔE between the presented and selected shade was calculated for each participant under the two lighting conditions. The significance was statistically assessed using a paired t test. RESULTS: The average ΔE between presented and selected shade for individual participants under natural daylight ranged from 0 to 4.84, with a mean of 2.24, while those under daylight lamp conditions ranged from 0 to 3.68, with a mean of 1.14. The difference was statistically significant, with P < 0.0001. CONCLUSION: Daylight lamp conditions significantly improved the shade matching performance of dental students.