KRAS-mutant non-small cell lung cancer (NSCLC) therapy based on tepotinib and omeprazole combination.

BACKGROUND: KRAS-mutant non-small cell lung cancer (NSCLC) shows a relatively low response rate to chemotherapy, immunotherapy and KRAS-G12C selective inhibitors, leading to short median progression-free survival, and overall survival. The MET receptor tyrosine kinase (c-MET), the cognate receptor of hepatocyte growth factor (HGF), was reported to be overexpressed in KRAS-mutant lung cancer cells leading to tumor-growth in anchorage-independent conditions. METHODS: Cell viability assay and synergy analysis were carried out in native, sotorasib and trametinib-resistant KRAS-mutant NSCLC cell lines. Colony formation assays and Western blot analysis were also performed. RNA isolation from tumors of KRAS-mutant NSCLC patients was performed and KRAS and MET mRNA expression was determined by real-time RT-qPCR. In vivo studies were conducted in NSCLC (NCI-H358) cell-derived tumor xenograft model. RESULTS: Our research has shown promising activity of omeprazole, a V-ATPase-driven proton pump inhibitor with potential anti-cancer properties, in combination with the MET inhibitor tepotinib in KRAS-mutant G12C and non-G12C NSCLC cell lines, as well as in G12C inhibitor (AMG510, sotorasib) and MEK inhibitor (trametinib)-resistant cell lines. Moreover, in a xenograft mouse model, combination of omeprazole plus tepotinib caused tumor growth regression. We observed that the combination of these two drugs downregulates phosphorylation of the glycolytic enzyme enolase 1 (ENO1) and the low-density lipoprotein receptor-related protein (LRP) 5/6 in the H358 KRAS G12C cell line, but not in the H358 sotorasib resistant, indicating that the effect of the combination could be independent of ENO1. In addition, we examined the probability of recurrence-free survival and overall survival in 40 early lung adenocarcinoma patients with KRAS G12C mutation stratified by KRAS and MET mRNA levels. Significant differences were observed in recurrence-free survival according to high levels of KRAS mRNA expression. Hazard ratio (HR) of recurrence-free survival was 7.291 (p = 0.014) for high levels of KRAS mRNA expression and 3.742 (p = 0.052) for high MET mRNA expression. CONCLUSIONS: We posit that the combination of the V-ATPase inhibitor omeprazole plus tepotinib warrants further assessment in KRAS-mutant G12C and non G12C cell lines, including those resistant to the covalent KRAS G12C inhibitors.

Dietary interventions and nutritional impact on oral health and development: a review.

Food and nutrients play a pivotal role in the development of dentition and oral health. While diet includes the overall forms of consumed foods, the nutrients include specific micro- (vitamins and minerals) and macro- (carbohydrates, proteins, and lipids) nutrients. As much as the assimilation of macro and micronutrients depends on proper oral health for ingesting food, oral health, in turn, depends on the availability of nutrients in the food so ingested. Age, certain medical conditions, socioeconomic status, and overall changes in the evolution of society determine the type of diet which in turn influences oral health. This article has touched upon certain salient aspects of these nutrients and their bearing on overall oral health and development.

BAHD acetyltransferase contributes to wound-induced biosynthesis of oleo-gum resin triterpenes in Boswellia.

Triterpenes (30-carbon isoprene compounds) represent a large and highly diverse class of natural products that play various physiological functions in plants. The triterpene biosynthetic enzymes, particularly those catalyzing the late-stage regio-selective modifications are not well characterized. The bark of select Boswellia trees, e.g., B. serrata exudes specialized oleo-gum resin in response to wounding, which is enriched with boswellic acids (BAs), a unique class of C3α-epimeric pentacyclic triterpenes with medicinal properties. The bark possesses a network of resin secretory structures comprised of vertical and horizontal resin canals, and amount of BAs in bark increases considerably in response to wounding. To investigate BA biosynthetic enzymes, we conducted tissue-specific transcriptome profiling and identified a wound-responsive BAHD acetyltransferase (BsAT1) of B. serrata catalyzing the late-stage C3α-O-acetylation reactions in the BA biosynthetic pathway. BsAT1 catalyzed C3α-O-acetylation of αBA, ßBA, and 11-keto-ßBA in vitro and in planta assays to produce all the major C3α-O-acetyl-BAs (3-acetyl-αBA, 3-acetyl-ßBA, and 3-acetyl-11-keto-ßBA) found in B. serrata bark and oleo-gum resin. BsAT1 showed strict specificity for BA scaffold, whereas it did not acetylate the more common C3ß-epimeric pentacyclic triterpenes. The analysis of steady-state kinetics using various BAs revealed distinct substrate affinity and catalytic efficiency. BsAT1 transcript expression coincides with increased levels of C3α-O-acetyl-BAs in bark in response to wounding, suggesting a role of BsAT1 in wound-induced biosynthesis of C3α-O-acetyl-BAs. Overall, the results provide new insights into the biosynthesis of principal chemical constituents of Boswellia oleo-gum resin.

Acute-onset Restless legs syndrome in acute neurological conditions-a prospective study on patients with the Guillain-Barre syndrome and acute stroke.

OBJECTIVES: While the Restless legs syndrome (RLS) is usually recognized as a chronic condition, it has often been diagnosed among patients with acute neurological illnesses, in which limb discomfort is reported. This study was conducted to determine how many among these, actually have acute-onset RLS, and also to evaluate characteristics of this subgroup of patients with Guillain-Barre syndrome (GBS) and stroke developing acute-onset RLS. METHODS: Consecutive patients diagnosed with GBS and eligible stroke patients, admitted to our Neurology services over a 1-year period, were enrolled. They were evaluated for symptoms of RLS based on IRLSSG consensus criteria and the AIIMS RLS Questionnaire for Indian patients (ARQIP). RESULTS: Forty adults with GBS and 58 with stroke were included. A total of 10 of the 40 (25%) patients with GBS developed definite acute RLS, which was mostly monophasic. Seven (70%) of these had demyelinating type of GBS, a significant association with acute RLS (P = .024). Six of the 58 stroke patients (10%) developed definite acute-onset, often persistent RLS. Subcortical location showed significant association with increased risk of developing acute RLS (P < .001). All patients diagnosed with acute-onset RLS had an immediate and good response to dopamine agonists. CONCLUSION: This is the first study showing that acute-onset RLS is common, affecting nearly 25% of patients with GBS and 10% patients with acute stroke. Recognizing and treating it can majorly contribute toward symptom relief and early improvement in the quality of life for this population.

Plant growth-promoting rhizobacteria enhance wheat salt and drought stress tolerance by altering endogenous phytohormone levels and TaCTR1/TaDREB2 expression.

Abiotic stresses such as salt and drought represent adverse environmental conditions that significantly damage plant growth and agricultural productivity. In this study, the mechanism of the plant growth-promoting rhizo-bacteria (PGPR)-stimulated tolerance against abiotic stresses has been explored. Results suggest that PGPR strains, Arthrobacter protophormiae (SA3) and Dietzia natronolimnaea (STR1), can facilitate salt stress tolerance in wheat crop, while Bacillus subtilis (LDR2) can provide tolerance against drought stress in wheat. These PGPR strains enhance photosynthetic efficiency under salt and drought stress conditions. Moreover, all three PGPR strains increase indole-3-acetic acid (IAA) content of wheat under salt and drought stress conditions. The SA3 and LDR2 inoculations counteracted the increase of abscisic acid (ABA) and 1-aminocyclopropane-1-carboxylate (ACC) under both salt and drought stress conditions, whereas STR1 had no significant impact on the ABA and ACC content. The impact of PGPR inoculations on these physiological parameters were further confirmed by gene expression analysis as we observed enhanced levels of the TaCTR1 gene in SA3-, STR1- and LDR2-treated wheat seedlings as compared to uninoculated drought and salt stressed plants. PGPR inoculations enhanced expression of TaDREB2 gene encoding for a transcription factor, which has been shown to be important for improving the tolerance of plants to abiotic stress conditions. Our study suggest that PGPR confer abiotic stress tolerance in wheat by enhancing IAA content, reducing ABA/ACC content, modulating expression of a regulatory component (CTR1) of ethylene signaling pathway and DREB2 transcription factor.

Alpha hemoglobin stabilizing protein: Its causal relationship with the severity of beta thalassemia.

Thalassemia major is characterized by anemia, iron overload and cellular damage. The severity of symptoms correlates with the alpha/non-alpha globin imbalance and is proportional to the magnitude of alpha chain excess. Alpha hemoglobin stabilizing protein (AHSP), the erythroid specific alpha globin chaperone, stabilizes free alpha chains, and prevents the formation of reactive oxygen radicals. Though AHSP expression has been linked to the severity of beta thalassemia, its role as a probable genetic modifier of disease severity, has still not been unequivocally established. In the present study, the level of the chaperone has been seen to vary in regularly transfused beta thalassemia patients, being underexpressed in 64% of cases, upregulated in 16% and comparable to controls in 20% of the cases. This discrepancy may be attributed to the degree of DNA damage, % HbF, and the number of nucleated RBCs in the peripheral blood of these patients. Results reveal that a decrease in the free alpha chain pool, and hence the repertoire of unbound iron, due to elevated HbF and/or the presence of nucleated RBCs in the peripheral blood results in the upregulation of the AHSP gene.

DNA damage: beta zero versus beta plus thalassemia.

BACKGROUND: ß thalassemia results in an increase in the α to non-α chain ratio. Iron released from unpaired α chains in RBCs and that ensuing from regular transfusions is the major cause of cellular damage. The use of iron chelators to counter the iron overload is accompanied by side-effects. The extent of iron toxicity could vary from one patient to another and could help in determining the optimal chelator dose for each patient. AIM: To observe the pro-oxidant/antioxidant disturbance and the extent of DNA damage in ß thalassemia patients with different ß globin gene anomalies. METHODS: The formation of Reactive Oxygen Species (ROS ) was observed by incubation of cell suspensions with 2',7', dichlorofluorescin-diacetate (DCFH DA) and DNA damage was demonstrated by single cell gel electrophoresis. Heinz bodies were observed by staining blood smears. SUBJECTS: The study group comprised 50 regularly transfused beta thalassemia patients and 40 non thalassemic controls. RESULTS: While Heinz bodies and nucleated RBCs were seen in all the patients, oxidation of DCFH and DNA damage were seen to be associated with the ß globin gene defect. DNA damage was found to be greater in ß(0) homozygotes as compared to the ß(+) homozygotes, and was maximum in patients presenting with the 619 base pair deletion. CONCLUSION: In the present study, iron toxicity, as indicated by DNA damage, has been seen to vary in the patients. Thus, monitoring of the dose of iron chelators, according to the type of mutation in the beta globin gene, may help improve the compliance of beta thalassemics to chelation therapy and prevent side-effects in patients with beta plus mutations.

4-coumarate: CoA ligase partitions metabolites for eugenol biosynthesis.

Biosynthesis of eugenol shares its initial steps with that of lignin, involving conversion of hydroxycinnamic acids to their corresponding coenzyme A (CoA) esters by 4-coumarate:CoA ligases (4CLs). In this investigation, a 4CL (OS4CL) was identified from glandular trichome-rich tissue of Ocimum sanctum with high sequence similarity to an isoform (OB4CL_ctg4) from Ocimum basilicum. The levels of OS4CL and OB4CL_ctg4-like transcripts were highest in O. sanctum trichome, followed by leaf, stem and root. The eugenol content in leaf essential oil was positively correlated with the expression of OS4CL in the leaf at different developmental stages. Recombinant OS4CL showed the highest activity with p-coumaric acid, followed by ferulic, caffeic and trans-cinnamic acids. Transient RNA interference (RNAi) suppression of OS4CL in O. sanctum leaves caused a reduction in leaf eugenol content and trichome transcript level, with a considerable increase in endogenous p-coumaric, ferulic, trans-cinnamic and caffeic acids. A significant reduction in the expression levels was observed for OB4CL_ctg4-related transcripts in suppressed trichome compared with transcripts similar to the other four isoforms (OB4CL_ctg1, 2, 3 and 5). Sinapic acid and lignin content were also unaffected in RNAi suppressed leaf samples. Transient expression of OS4CL-green fluorescent protein fusion protein in Arabidopsis protoplasts was associated with the cytosol. These results indicate metabolite channeling of intermediates towards eugenol by a specific 4CL and is the first report demonstrating the involvement of 4CL in creation of virtual compartments through substrate utilization and committing metabolites for eugenol biosynthesis at an early stage of the pathway.

Novel urea and thiourea derivatives of thiazole-glutamic acid conjugate as potential inhibitors of microbes and fungi.

Since discovery and development of effective as well as safe drugs has brought a progressive era in human healthcare that is accompanied by the appearance of drug resistant bacterial strains, there is constant need of new antibacterial agent having novel mechanisms of action to act against the harmful pathogens. In the present study, several N-terminal substituted urea/thiourea derivatives were synthesized by the reaction of glutamic acid and 3-(1-piperazinyl)-1,2-benzisothiazole with various substituted phenyl isocyanates/isothiocyanates. Elemental analysis, IR, 1H NMR, 13C NMR and mass spectral data confirmed the structure of the newly synthesized compounds. The derivatives were investigated for their antibacterial and antifungal activities against various pathogens of human origin by agar well diffusion method and microdilution method. The preliminary antimicrobial bioassay reveals that the compounds containing fluoro and bromo as substituents showed promising antimicrobial activity.

Relationship between femoral component placement and patient-specific anatomical rotational landmarks in robotic arm assisted total knee Arthroplasty- a multicentric study.

Introduction: Osteoarthritis of the knee is a common problem in the elderly, leading to severe morbidity. Total Knee Arthroplasty (TKA) is a widely validated surgery to provide a remarkable extent of knee function and simultaneously alleviates pain for knee osteoarthritis (OA). It is clearly understood that precision of the rotational alignment and accuracy of the technique in the placement of the femoral component is a prerequisite for excellent and successful outcomes of TKA. Advanced technology has now allowed surgeons to understand patient-specific variabilities in anatomical reference landmarks and the relationship of component positioning in relation to the reference landmarks to achieve accurate gap balancing with minimal soft tissue release.14 Robotic Arm Assisted-Total Knee Arthroplasty (RAA-TKA) is a semi-automated system that enables us in replicating the same. Using this technology, the bony resections, component positions, probable component sizing and gap balancing can be tentatively planned preoperatively with CT Scan Analysis and executed intraoperatively. Hence this study was undertaken to estimate the relationship between femoral component placement to normal rotational landmarks such as the Posterior Condylar Axis (PCA) and to quantitatively evaluate coronal and sagittal plane correction obtained. Also, we aimed to use the data to detect any anatomical variations in the study population and evaluate the accuracy of predicted component sizing, including gender-based evaluation. Materials and methods: A Prospective Observational Study of 1073 knees of patients of either sex above 50 years of age with Kellgren Lawrence Grade 4 Osteoarthritis of the knee which were confirmed with X-Ray undergoing RAA-TKA using MAKO Robotic System using Stryker Triathlon (Cruciate Substituting) CS Knee was conducted during the period between 2022 and 2023 in two South Indian hospitals specializing in joint replacement surgeries. Results: We found a statistically significant difference between the native Posterior Condylar Axis (PCA) (4.82 ± 2.15°) and final femoral component external rotation (3.24 ± 1.29°) with a p-value of <0.001 at 95% confidence interval. The accuracy of component size prediction was 99.8%. Also, analysis in our study has shown the most common implant sizes to be 4 in males and 2 in females. We also found no statistically significant difference based on age, size, laterality, or primary varus deformity. Conclusions: RAA-TKA provides patient-specific alignment/restricted kinematic alignment which might further enhance the outcome for the patient. Reliable deformity correction in coronal and sagittal planes can be achieved. Accurate flexion and extension gap balancing can be done through component placement and with minimal soft tissue dissection. Irrespective of all the advantages noted in RAA-TKA, further follow-up and long-term outcome studies are required to properly gauge and analyze this new technology.

Genetically targeted chemical assembly of polymers specifically localized extracellularly to surface membranes of living neurons.

Multicellular biological systems, particularly living neural networks, exhibit highly complex organization properties that pose difficulties for building cell-specific biocompatible interfaces. We previously developed an approach to genetically program cells to assemble structures that modify electrical properties of neurons in situ, opening up the possibility of building minimally invasive cell-specific structures and interfaces. However, the efficiency and biocompatibility of this approach were challenged by limited membrane targeting of the constructed materials. Here, we design a method for highly localized expression of enzymes targeted to the plasma membrane of primary neurons, with minimal intracellular retention. Next, we show that polymers synthesized in situ by this approach form dense extracellular clusters selectively on the targeted cell membrane and that neurons remain viable after polymerization. Last, we show generalizability of this method across a range of design strategies. This platform can be readily extended to incorporate a broad diversity of materials onto specific cell membranes within tissues and may further enable next-generation biological interfaces.

Inferior alveolar nerve dysfunction in mandibular fractures: a prospective cohort study.

OBJECTIVES: To assess the prevalence and recovery of inferior alveolar nerve dysfunction (IAND) in mandibular fractures. MATERIALS AND METHODS: : This was a prospective cohort study. Clinical neurosensory testing was done preoperatively and the IAND was categorized as mild, moderate or severe. Postoperatively, neurosensory testing was repeated at 1 day, 1 week, 1 month, 3 months and every 3 months thereafter. RESULTS: : A total of 257 patients with 420 fractures were included in the study with a mean age of 31.7 years. Body fractures (95.9%) had the highest incidence of IAND, followed by the angle fractures (90.1%) and symphysis fractures (27.6%). The condyle and coronoid fractures did not have any IAND and hence were excluded from further study. After eliminating those cases, 232 patients remained in the study with 293 fractures. The overall prevalence of IAND in fractures occurring distal to the mandibular foramen was 56.3%. The changes until 1 week were minimal. From 1 month to 6 months, there was a significant reduction in the severity of IAND. A significant number of cases (60.0%) were lost to follow-up between 6 and 9 months. At 6 months, 23.9% of cases still had some form of IAND and 95.0% of the symphysis, 59.0% of the angle and 34.8% of the body fractures with IAND had become normal. CONCLUSION: This study documents the reduction in the degree of severity of IAND in the first six months and provides the basis for future studies with longer periods of follow-up.

Chemical diversity in the genus Alpinia (Zingiberaceae): comparative composition of four Alpinia species grown in Northern India.

The essential-oil compositions of leaves, flowers, and rhizomes of Alpinia galanga (L.) Willd., Alpinia calcarata Rosc., Alpinia speciosa K. Schum., and Alpinia allughas Rosc. were examined and compared by capillary GC and GC/MS. Monoterpenoids were the major oil constituents identified. 1,8-Cineole, alpha-terpineol, (E)-methyl cinnamate, camphor, terpinen-4-ol, and alpha- and beta-pinenes were the major constituents commonly distributed in leaf and flower essential oils. The presence of endo-fenchyl acetate, exo-fenchyl acetate, and endo-fenchol was the unique feature of rhizome essential oils of A. galanga, A. calcarata, and A. speciosa. On contrary, the rhizome oil of A. allughas was dominated by beta-pinene. Significant qualitative and quantitative variations were observed in essential-oil compositions of different parts of Alpinia species growing in subtemperate and subtropical regions of Northern India. Cluster analysis was performed to find similarities and differences in essential-oil compositions based on representative molecular skeletons. Monoterpenoids, viz., 1,8-cineole, terpinen-4-ol, camphor, pinenes, (E)-methyl cinnamate, and fenchyl derivatives, were used as chemotaxonomic markers.

Primary Malignant Melanoma of Oral Mucosa - Report of Two Cases.

Primary malignant melanoma of oral mucosa is a rare and aggressive tumor. It is usually seen in the 5th and 6th decades of life. Its mainstay of treatment is surgery. It has a very poor prognosis, which is attributed to its late detection and distant metastasis. Dentists are often the first clinicians to come across these lesions and need to be able to identify them at the earliest for a better prognosis. In this article, we present two cases of extensive primary malignant melanoma of the oral cavity. Clinically, both the cases had a similar appearance of grayish-black pigmented nodular swelling on the buccal aspect and grayish-black discoloration on the palatal aspect. There were no significant radiological changes in both cases, indicating the superficial spread of the lesion. A positron emission tomography scan was performed in the second patient, which did not show any distant metastasis. Surgery was advised as a treatment for both the patients.

Assessment of the Need for Routine Distolingual Local Anesthetic Infiltration in Addition to Traditional Inferior Alveolar, Lingual and Long Buccal Nerve Blocks in Mandibular Third Molar Extractions.

BACKGROUND AND OBJECTIVES: Persistent pain during the removal of mandibular third molars is often due to accessory nerve supply causing inadequate local anesthesia. This study aims to assess the requirement of routine distolingual infiltration anesthesia in addition to traditional inferior alveolar, lingual, and long buccal nerve block in mandibular third molar extractions. METHODOLOGY: Sixty patients requiring mandibular third molar extraction were randomly divided into two equal groups; Group A (Classic inferior alveolar, lingual, and buccal nerve block) and Group B (with an additional 0.2 ml distolingual infiltration). During various steps of the procedure, any complaint of pain was recorded and graded on a subjective Visual Analog Scale (VAS). RESULTS: There was no significant difference between the two groups in regard to age (P = 0.666) and sex (P = 0.432). And also, no difference was found in angulation (P = 0.757), class (P = 0.417) and position (P = 1.000) of third molars. Mean VAS scores in Group B (0.153) were significantly lower (P = 0.004) than that of Group A (0.600). VAS scores during procedural steps were significantly lower in Group B during mucoperiosteal elevation (P = 0.050), bone guttering (P = 0.037), and tooth splitting (P = 0.052). CONCLUSION: Routine distolingual infiltration anesthesia, in addition to classic inferior alveolar, lingual, and long buccal nerve block, is recommended for the extraction of mandibular third molars.

Assessment of Healing Using Alvogyl and Platelet Rich Fibrin in Patients with Dry Socket - An Evaluative Study.

INTRODUCTION: Dry socket is one of the most common postoperative complications following the extraction of permanent teeth, which is characterized by pain and exposed bone. The usual protocol followed for its management is irrigation of the socket and packing of the socket with medicated gel or paste to provide relatively faster pain relief and allow normal wound healing. In this study, we evaluated the outcome of management of dry socket with platelet-rich fibrin (PRF) and intraalveolar alvogyl dressing, in terms of improvement in pain and socket epithelialization after the treatment. METHODOLOGY: Thirty participants with established dry socket were randomly divided into two groups: Group A and Group B. The participants in Group A were treated with alvogyl and those in Group B were treated with PRF. Clinical parameters were assessed for both groups on the 1st day of the procedure and on the 3rd and 10th-day postoperatively for the reduction in pain and wound healing. RESULTS: There was a significant decrease in pain and the number of socket wall exposure in both the groups by the 3rd postoperative day. In both the groups, the pain had completely resolved and socket fully epithelialized by the 10th postoperative day. DISCUSSION: The use of PRF in the present study yielded promising results in terms of both pain reduction and improved wound healing which was comparable to the conventional alvogyl dressing. It may be concluded that PRF is an effective modality for the management of dry socket.

Induced mutation breeding for qualitative and quantitative traits and varietal development in medicinal and aromatic crops at CSIR-CIMAP, Lucknow (India): past and recent accomplishment.

PURPOSE: The nature and kind of some successful mutations achieved through gamma radiation at CSIR-CIMAP, Lucknow are reviewed and described in this article. Medicinal and aromatic crops (MACs) came under the mutation-breeding program very late - perhaps during the 1980s in India. Nevertheless, successes have been glaring and unique too. The plant breeders of the Institute concerted attempts were made for the genetic restructuring of the plant-frame in Hyoscyamus niger L. (black henbane) and Cymbopogon martinii (palmarosa), enhanced alkaloid biosynthesis in henbane, weak, or lack of latex biosynthesis in Papaver somniferum L. (Opium poppy), increased productivity of seeds in Plantago ovata F. (isabgol), and new/increased formation of essential oils in Chamomila recutita [L.] Rauschert (Chamomile) and Cymbopogon winterianus (Java citronella). Several qualitative macro mutations were generated for commercial exploitation, e.g. an unbranched (Ub) and physiologically most efficient variety Aekla and high tropane alkaloid bearing variety Aela of H. niger, opium less oil-seed variety Sujata of opium poppy, a dwarf mutant and a male sterile (ms) line of palmarosa, and new compound yielding variety of chamomile and geranium. CONCLUSIONS: The ample quantitative variation was also created by reshuffling the polygenic background in both seed and vegetatively propagated MACs, and subsequently, with applying mutation breeding approach superior varieties were evolved and released after a rigorous screening in the field-evaluation or under pipeline for release. These varieties are Niharika and Mayuri of psyllium; Vallary, CIMAP Sammohak, CIM Ujjwala, and YEL (yellow) of German chamomile and Manjari and CIM Jeeva of Java citronella.