Fabrication of three-dimensionally printed polylactic acid nasal stent prosthesis for postnasal reconstruction using extraoral scanning and photogrammetry techniques: A report on two patients.

Severe and combined nasal defects associated with trauma or neoplasm excision can cause significant functional and esthetic problems. To avoid nasal synechia following reconstructive surgeries, a nasal stent prosthesis is required to act as an internal scaffold to support the graft and residual tissues. The purpose of the stent is to maintain internal airway patency and to prevent collapse and contracture of the donor tissues. A conventional nasal stent prosthesis has disadvantages, including the difficulty in maintaining adequate thickness and internal patency during fabrication. Hence, this clinical report introduces the fabrication technique for 3- dimensionally printed polylactic acid nasal stent prostheses using extraoral scanning and photogrammetry methods.

Rehabilitation Using Hybrid Technique for Feeding Plate Fabrication in a 3-Month-Old Infant with Cleft Palate Defect Utilising Digital and Conventional Method: A Case Report.

ABSTRACT: Cleft palate leads to difficulty in suckling, speech abnormalities, dental problems, hearing loss and middle ear infections. Feeding plate acts as a barrier between the oral and nasal cavities allowing the normal development of jaws and enhanced nutritional supply to the patient. Fabrication of the feeding plate using digital means provide better adaptability due to the engagement of all the possible favourable anatomic undercuts, thereby, providing better retention. This case report discusses the hybrid technique including both the digital and conventional means for fabrication of a feeding plate to obturate the defect between the oral and nasal cavity.

Validation of candidate gene-based EST-SSR markers for sugar yield in sugarcane.

Sugarcane (Saccharum spp.) is a widely cultivated crop that fulfils approximately 75% of the sucrose demand worldwide. Owing to its polyploidy and complex genetic nature, it is difficult to identify and map genes related to complex traits, such as sucrose content. However, association mapping is one of the alternatives for identifying genes or markers for marker-assisted selection. In the present study, EST-SSR primers were obtained from in silico studies. The functionality of each primer was tested using Blast2Go software, and 30 EST-SSR primers related to sugar content were selected. These markers were validated using association analysis. A total of 70 F1 diverse genotypes for sugar content were phenotypes with two check lines. All parameters related to sugar content were recorded. The results showed a significant variation between the genotypes for sugar yield traits such as Brix value, purity, and sucrose content, etc. Correlation studies revealed that the Brix%, sucrose content, and sucrose recovery were significantly correlated. An association analysis was performed using mixed linear model to avoid false positive associations. The association analysis revealed that the SEM 407 marker was significantly associated with Brix% and sucrose content. The SEM 407 primers are putatively related to diphosphate-fructose-6-phosphate 1-phosphotransferase which is associated with Brix% and sucrose content. This functional marker can be used for marker-assisted selection for sugar yield traits in sugarcane that could accelerate the sugarcane breeding program.

Magnet-Retained closed bulb hollow obturator and orbital prosthesis for patient with maxillectomy and orbital exenteration following COVID-Associated mucormycosis.

The rehabilitation of facial deformities is a challenging endeavour that necessitates customising the procedure for each patient. Significant physical and psychological impacts might arise as a result of the deformity in the orofacial region. Post-COVID rhino-orbital mucormycosis has led to rise in extraoral and intraoral defects since 2020. To avoid further surgery, an economical maxillofacial prosthesis is an excellent choice as it is aesthetic, durable, long-lasting and retentive. This case report describes the prosthetic rehabilitation of the patient with post-COVID mucormycosis maxillectomy and orbital exenteration using a magnet-retained closed bulb hollow acrylic obturator and room-temperature vulcanising silicone orbital prosthesis. To enhance retention, a spectacle and medical-grade adhesive were also used.

Surgical Rehabilitation of a Continuous Orbital and Maxillary Defect from Rhino-Orbital Mucormycosis Utilising Digital Technology - A Case Report.

Rationale: COVID-19 has led to a resurgence in cases of mucormycosis, especially the rhino-orbital form affecting the oral cavity, nasal, orbital and cerebral regions. Patient Concerns: The surgical treatment in this patient led to the exenteration of orbital contents and segmental maxillectomy of the affected side leading to facial disfigurement and inability to masticate. Diagnosis: A combined mucormycosis-associated oro-orbital defect was present leading to a communication between oral and orbital cavities. Treatment: Rehabilitation utilising digital technology for removable prosthesis was planned for the combined orbital and oral defect. Outcomes: The independent intraoral and orbital prosthesis reduced the mobility of the orbital prosthesis while performing functional movements. The advancements in digital technology led to the convenient and resilient fabrication of prostheses for large facial defects. Take-away Lessons: The prosthetic rehabilitation of a continuous orbital and oral defect with a hybrid of both digital and conventional means provided an aesthetic, feasible and financially sound solution to the patient.

mTORC1 functional assay reveals SZT2 loss-of-function variants and a founder in-frame deletion.

Biallelic pathogenic variants in SZT2 result in a neurodevelopmental disorder with shared features, including early-onset epilepsy, developmental delay, macrocephaly, and corpus callosum abnormalities. SZT2 is as a critical scaffolding protein in the amino acid sensing arm of the mTORC1 signalling pathway. Due to its large size (3432 amino acids), lack of crystal structure, and absence of functional domains, it is difficult to determine the pathogenicity of SZT2 missense and in-frame deletions, but these variants are increasingly detected and reported by clinical genetic testing in individuals with epilepsy. To exemplify this latter point, here we describe a cohort of 12 individuals with biallelic SZT2 variants and phenotypic overlap with SZT2-related neurodevelopmental disorders. However, the majority of individuals carried one or more SZT2 variants of uncertain significance (VUS), highlighting the need for functional characterization to determine, which, if any, of these VUS were pathogenic. Thus, we developed a novel individualized platform to identify SZT2 loss-of-function variants in the context of mTORC1 signalling and reclassify VUS. Using this platform, we identified a recurrent in-frame deletion (SZT2 p.Val1984del) which was determined to be a loss-of-function variant and therefore likely pathogenic. Haplotype analysis revealed that this single in-frame deletion is a founder variant in those of Ashkenazi Jewish ancestry. Moreover, this approach allowed us to tentatively reclassify all of the VUS in our cohort of 12 individuals, identifying five individuals with biallelic pathogenic or likely pathogenic variants. Clinical features of these five individuals consisted of early-onset seizures (median 24 months), focal seizures, developmental delay and macrocephaly similar to previous reports. However, we also show a widening of the phenotypic spectrum, as none of the five individuals had corpus callosum abnormalities, in contrast to previous reports. Overall, we present a rapid assay to resolve VUS in SZT2, identify a founder variant in individuals of Ashkenazi Jewish ancestry, and demonstrate that corpus callosum abnormalities is not a hallmark feature of this condition. Our approach is widely applicable to other mTORopathies including the most common causes of the focal genetic epilepsies, DEPDC5, TSC1/2, MTOR and NPRL2/3.

Plasma Membrane Receptors Involved in the Binding and Response of Osteoclasts to Noncellular Components of the Bone.

Osteoclasts differentiate from hematopoietic cells and resorb the bone in response to various signals, some of which are received directly from noncellular elements of the bone. In vitro, adherence to the bone triggers the reduction of cell-cell fusion events between osteoclasts and the activation of osteoclasts to form unusual dynamic cytoskeletal and membrane structures that are required for degrading the bone. Integrins on the surface of osteoclasts are known to receive regulatory signals from the bone matrix. Regulation of the availability of these signals is accomplished by enzymatic alterations of the bone matrix by protease activity and phosphorylation/dephosphorylation events. Other membrane receptors are present in osteoclasts and may interact with as yet unidentified signals in the bone. Bone mineral has been shown to have regulatory effects on osteoclasts, and osteoclast activity is also directly modulated by mechanical stress. As understanding of how osteoclasts and other bone cells interact with the bone has emerged, increasingly sophisticated efforts have been made to create bone biomimetics that reproduce both the structural properties of the bone and the bone's ability to regulate osteoclasts and other bone cells. A more complete understanding of the interactions between osteoclasts and the bone may lead to new strategies for the treatment of bone diseases and the production of bone biomimetics to repair defects.

Mitochondrial Dysfunction in Fragile-X Syndrome: Plugging the Leak May Save the Ship.

Recent work by Licznerski et al. suggests that mutant FMRP linked to Fragile-X syndrome elevates the inner mitochondrial membrane proton leak, leading to increased metabolism and changes in protein synthesis that trigger impaired synaptic maturation and autistic behaviors.

Ropinirole, a potential drug for systematic repositioning based on side effect profile for management and treatment of breast cancer.

Drug repositioning offers two main advantages in drug discovery - the process is less tedious and less costly. In the past, many drugs like thalidomide and sildenafil were successfully repositioned but the process was entirely serendipitous. These days drug repositioning is widely accepted as an alternate method of drug discovery and the process is based on building a strong hypothesis guided by systematic computational and experimental methods. One of the methods used in drug repositioning is based on shared side effects by drugs of different pharmacological categories. This method rests on the principle that drugs that share side effects might also share common biological targets and therefore same pharmacological indications. Old drugs can be repositioned for new uses by identifying the shared side effects of existing drugs and by modulating their chemical structure if required. Breast cancer is the most common type of cancer in women and the second leading cause of death worldwide after lung cancer in both men and women. Letrozole, an aromatase inhibitor, is used in the treatment of advanced, recurrent and metastatic breast cancer in post-menopausal women. Identification of drugs that share side effects with letrozole might help us to identify a potential drug for repositioning in the treatment of breast cancer. Ropinirole, a dopaminergic agonist was found to share the maximum number of side effects with letrozole. Studies have proposed that dopaminergic agonists induce apoptosis in breast, colon, ovarian cancer cells and leukemia neuroblastoma. This is consistent with our hypothesis that ropinirole that shares the maximum number of side effects with letrozole might be effective in the management of breast cancer. This hypothesis was further validated by preliminary molecular docking and in-vitro cell-line studies.

Iminoenamine based novel androgen receptor antagonist exhibited anti-prostate cancer activity in androgen independent prostate cancer cells through inhibition of AKT pathway.

Treatment by androgen receptor (AR) antagonists is one of the regimens for prostate cancer. The prolonged treatment with AR antagonist leads to the expression of point mutation in the ligand binding domain of the AR. This point mutation causes resistance to AR antagonist by converting them into an agonist. The T887A mutated AR was frequently expressed in androgen independent prostate cancer (AIPC) patients. Through literature survey and molecular modelling, we have identified a novel AR antagonist having a bulky ß-iminoenamine BF2 complex scaffold. The tested and standard ligands were screened in AR positive (LNCaP, MCF-7 and MDA-MB-453), AR negative (PC3), and non-cancerous (3T3) cell lines through anti-proliferation assay. The ligand, ARA3 was the most potent molecule among all the tested ligands and was 7.6 folds selective for AR positive cell lines. The mechanism of anti-prostate cancer activity of ARA3 was confirmed by western blot, qPCR, and apoptotic assays in LNCaP (T887A positive AR) cells. Structural activity relationship was derived by correlating the in-vitro and in-silico data. Consequently, we have identified the essential functional groups that could prevent the resistance concerning mutant AR. The ARA3 induces the apoptosis in AIPC cells by preventing the AR mediated activation of AKT pathway. The bicalutamide did not induce the apoptosis because it failed to prevent the AR mediated activation of AKT.

Design, synthesis and biological evaluation of 2-(4-phenylthiazol-2-yl) isoindoline-1,3-dione derivatives as anti-prostate cancer agents.

The structural modification and molecular docking-based screening approaches on thiazole-based isoindolinediones were imposed to find the novel 2-(4-phenylthiazol-2-yl) isoindoline-1,3-dione derivatives. The best fit compounds (6a-n) were synthesized and evaluated their antiproliferative activities on the prostate cancer cell lines (PC-3 & LNCaP). Among them, the compound, 6m exhibited good activity, particularly on LNCaP (IC50=5.96±1.6µM), moderately active against PC-3 cell lines as compared to bicalutamide. The compound, 6m decreased the androgen-mediated transcription of ARE-mRNA in PSA, TMPRSS2, c-myc and cyclin D1 than R-bicalutamide. The compounds, 6e and 6f were reconfirmed through single crystal XRD analysis. The ADME profiling of the test compounds was evaluated to find the drug-likeness and pharmacokinetic parameters. These findings may provide vital information for the development of anti-prostate cancer agents.

Dual inhibition of CDK4/Rb and PI3K/AKT/mTOR pathways by ON123300 induces synthetic lethality in mantle cell lymphomas.

This study describes the characterization of a novel kinase inhibitor, ON123300, which inhibits CDK4/6 (cyclin-dependent kinases 4 and 6) and phosphatidylinositol 3 kinase-δ (PI3K-δ) and exhibits potent activity against mantle cell lymphomas (MCLs) both in vitro and in vivo. We examined the effects of PD0332991 and ON123300 on cell cycle progression, modulation of the retinoblastoma (Rb) and PI3K/AKT pathways, and the induction of apoptosis in MCL cell lines and patient-derived samples. When Granta 519 and Z138C cells were incubated with PD0332991 and ON123300, both compounds were equally efficient in their ability to inhibit the phosphorylation of Rb family proteins. However, only ON123300 inhibited the phosphorylation of proteins associated with the PI3K/AKT pathway. Cells treated with PD0332991 rapidly accumulated in the G0/G1 phase of cell cycle as a function of increasing concentration. Although ON123300-treated cells arrested similarly at lower concentrations, higher concentrations resulted in the induction of apoptosis, which was not observed in PD0332991-treated samples. Mouse xenograft assays also showed a strong inhibition of MCL tumor growth in ON123300-treated animals. Finally, treatment of ibrutinib-sensitive and -resistant patient-derived MCLs with ON123300 also triggered apoptosis and inhibition of the Rb and PI3K/AKT pathways, suggesting that this compound might be an effective agent in MCL, including ibrutinib-resistant forms of the disease.

Localized CCR2 Activation in the Bone Marrow Niche Mobilizes Monocytes by Desensitizing CXCR4.

Inflammatory (classical) monocytes residing in the bone marrow must enter the bloodstream in order to combat microbe infection. These monocytes express high levels of CCR2, a chemokine receptor whose activation is required for them to exit the bone marrow. How CCR2 is locally activated in the bone marrow and how their activation promotes monocyte egress is not understood. Here, we have used double transgenic lines that can visualize CCR2 activation in vivo and show that its chemokine ligand CCL2 is acutely released by stromal cells in the bone marrow, which make direct contact with CCR2-expressing monocytes. These monocytes also express CXCR4, whose activation immobilizes cells in the bone marrow, and are in contact with stromal cells expressing CXCL12, the CXCR4 ligand. During the inflammatory response, CCL2 is released and activates the CCR2 on neighboring monocytes. We demonstrate that acutely isolated bone marrow cells co-express CCR2 and CXCR4, and CCR2 activation desensitizes CXCR4. Inhibiting CXCR4 by a specific receptor antagonist in mice causes CCR2-expressing cells to exit the bone marrow in absence of inflammatory insults. Taken together, these results suggest a novel mechanism whereby the local activation of CCR2 on monocytes in the bone marrow attenuates an anchoring signalling provided by CXCR4 expressed by the same cell and mobilizes the bone marrow monocyte to the blood stream. Our results also provide a generalizable model that cross-desensitization of chemokine receptors fine-tunes cell mobility by integrating multiple chemokine signals.

Impact of fiducial arrangement and registration sequence on target accuracy using a phantom frameless stereotactic navigation model.

Modern frameless stereotactic techniques utilize scalp fiducial markers for registration. Anecdotal reports from surgeons indicate a variety of methods for improving accuracy using different fiducial arrangements and registration sequences. The few published studies on registration accuracy do not provide a simple and systematic method for determining target accuracy. Nine different arrangements of ten fiducial markers were attached to a model. Ten separate markers were designated as targets for evaluation of registration accuracy. We systematically registered each of the arrangements over multiple trials, in one of four sequences, and then measured the targets. The target coordinates were compared against the established target values, and a root-mean-square deviation (RMSD) was derived. A systematic multivariate analysis determined the effects of different variables on the RMSD. We found no correlation between the "Registration Accuracy" provided by Medtronic (Medtronic Navigation, Louisville, CO, USA) and our RMSD representing targeting accuracy (R=0.008). RMSD did vary for different fiducial arrangements. We found no significant difference between the various sequences of fiducial arrangement. Thus, regardless of fiducial arrangement, registration sequence has no impact on accuracy. Fiducial arrangements distributed optimally across the skull, however, allowed for significantly improved accuracy. Further studies are required to determine which different arrangements of fiducials are relevant for specific procedures.

Intracellular mannose binding lectin mediates subcellular trafficking of HIV-1 gp120 in neurons.

Human immunodeficiency virus-1 (HIV-1) enters the brain early during infection and leads to severe neuronal damage and central nervous system impairment. HIV-1 envelope glycoprotein 120 (gp120), a neurotoxin, undergoes intracellular trafficking and transport across neurons; however mechanisms of gp120 trafficking in neurons are unclear. Our results show that mannose binding lectin (MBL) that binds to the N-linked mannose residues on gp120, participates in intravesicular packaging of gp120 in neuronal subcellular organelles and also in subcellular trafficking of these vesicles in neuronal cells. Perinuclear MBL:gp120 vesicular complexes were observed and MBL facilitated the subcellular trafficking of gp120 via the endoplasmic reticulum (ER) and Golgi vesicles. The functional carbohydrate recognition domain of MBL was required for perinuclear organization, distribution and subcellular trafficking of MBL:gp120 vesicular complexes. Nocodazole, an agent that depolymerizes the microtubule network, abolished the trafficking of MBL:gp120 vesicles, suggesting that these vesicular complexes were transported along the microtubule network. Live cell imaging confirmed the association of the MBL:gp120 complexes with dynamic subcellular vesicles that underwent trafficking in neuronal soma and along the neurites. Thus, our findings suggest that intracellular MBL mediates subcellular trafficking and transport of viral glycoproteins in a microtubule-dependent mechanism in the neurons.

SU-E-T-649: Evaluation of RapidArc- Based Stereotactic Cranial Radiotherapy Plans with MU Objective Using Multiple Non Coplanar Arcs in Comparison with Conventional Dynamic Conformal Arc Technique.

PURPOSE: Previous researches reported that RapidArc plans for stereotactic cranial radiotherapy have two to three times more MUs as compared to Conventional Dynamic Conformal Arc (DCA) Technique. This study aims to evaluate RapidArc plans using multiple non- coplanar arcs, developed with MU objective constraint in the optimization stage. METHODS: Five single brain metastasis and three multiple metastases cases previously planned using DCA techniques in BrainLab iPlan Version 4.1 were investigated in this study. For each case, the target was defined on CT-MR fused images in iPlan. The CT images and contours of these patients were exported from iPlan to Varian Eclipse TPS Version 8.6. For each case, a DCA plan and a RapidArc plan with multiple non-coplanar arcs with and without using MU objective in the optimization stage were generated using Varian Trilogy machine with Millennium 120 MLC keeping the same prescription and critical structure dose limits. All plans were evaluated according to Conformity Index (CI-modified Paddick) Homogeneity Index (HI), and the normal tissue volume receiving various dose levels (V80%, V50%, V25% and V10%). RESULTS: In all the plans, the target objectives were met and dose to OARs was within tolerance dose constraints. RapidArc plans with and without MU objective showed better CI and HI as supposed to DCA plans. V80%, V50%, V25% and V10% of normal tissue for RapidArc plans are equal or lesser than DCA plans. Single isocentre RapidArc plan for closely spaced multiple metastases cases showed better dose fall off between the lesions as supposed to DCA plans. RapidArc plans with MU objective resulted in comparable MUs as that of DCA plans. CONCLUSIONS: Our study showed RapidArc plans done with and without MU objective have no significant dosimetric difference in plan objectives. Therefore, multiple non-coplanar RapidArc plans with MU objective is clinically feasible and can provide better treatment plans than conventional DCA plans, especially for complicated cases.

Solid pseudopapillary tumour of pancreas: A rare neoplasm.

Solid pseudopapillary tumour (SPT) is the rare tumour of pancreas with unknown aetiology and good prognosis. Occurs predominantly in young women of reproductive age group. Not many cases have been reported from India. We report a case of a young woman who presented with persistent back pain having large tumour of body of pancreas treated successfully by left pancreatectomy.

Isolation of antioxidant compounds from the methanolic extract of the roots of Decalepis hamiltonii (Wight and Arn.).

The tuberous roots of Decalepis hamiltonii are consumed as pickles and beverages and are believed to possess health-promoting properties. We have investigated the antioxidant potential of the roots. The methanolic extract of the root showed a high antioxidant activity. The methanolic extract was fractionated on a silica gel column, which showed three major fractions with good antioxidant activity. The active fractions were further subjected to preparative thin layer and silica gel column chromatography, which yielded six pure compounds. The purified compounds were characterized by MS, 1H NMR, 13C NMR, and two-dimensional NMR spectroscopic techniques and identified as 2-hydroxy-4-methoxybenzaldehyde, p-anisaldehyde, vanillin, borneol, salicylaldehyde, and bis-2,3,4,6-galloyl-alpha/beta-D-glucopyranoside. The latter compound, named decalepin, is a new antioxidant molecule from the plant kingdom. The purified compounds showed antioxidant activities in in vitro assays such as inhibition of lipid peroxidation, hydroxyl radical, superoxide anion, and 1,1-diphenyl-2picrylhydrazyl radical scavenging. This is the first report of the antioxidant constituents of the roots of Decalepis hamiltonii.