A Noncanonical CD56dimCD16dim/- NK Cell Subset Indicative of Prior Cytotoxic Activity Is Elevated in Patients with Autoantibody-Mediated Neurologic Diseases.

Neuromyelitis optica spectrum disorder (NMOSD), myelin oligodendrocyte glycoprotein Ab disease, and autoimmune myasthenia gravis (MG) are autoantibody-mediated neurologic conditions where autoantibodies can induce Ab-dependent cellular cytotoxicity (ADCC), a NK cell-mediated effector function. However, whether ADCC is a pathogenic mechanism in patients with these conditions has not been confirmed. We sought to characterize circulatory NK cells using functional assays, phenotyping, and transcriptomics to elucidate their role in pathology. NK cells from NMOSD patients and MG patients with elevated disease burden exhibited reduced ADCC and CD56dimCD16hi NK cells, along with an elevated frequency of CD56dimCD16dim/- NK cells. We determined that ADCC induces a similar phenotypic shift in vitro. Bulk RNA sequencing distinguished the CD56dimCD16dim/- population from the canonical CD56dimCD16hi cytotoxic and CD56hiCD16- immunomodulatory subsets, as well as CD56hiCD16+ NK cells. Multiparameter immunophenotyping of NK cell markers, functional proteins, and receptors similarly showed that the CD56dimCD16dim/- subset exhibits a unique profile while still maintaining expression of characteristic NK markers CD56, CD94, and NKp44. Notably, expression of perforin and granzyme is reduced in comparison with CD56dimCD16hi NK cells. Moreover, they exhibit elevated trogocytosis capability, HLA-DR expression, and many chemokine receptors, including CCR7. In contrast with NMOSD and MG, myelin oligodendrocyte glycoprotein Ab disease NK cells did not exhibit functional, phenotypic, or transcriptomic perturbations. In summary, CD56dimCD16dim/- NK cells are a distinct peripheral blood immune cell population in humans elevated upon prior cytotoxic activity by the CD56dimCD16hi NK cell subset. The elevation of this subset in NMOSD and MG patients suggests prior ADCC activity.

Combined Effects of Glutaraldehyde-based Desensitizer and Nd: YAG Laser on Dentinal Tubules Occlusion.

AIM: The aim of this study was to assess the impact of Nd:YAG laser, glutaraldehyde-based desensitizer (GD), or their combination on occluding dentinal tubules. MATERIALS AND METHODS: Fifty dentin samples were obtained from non-carious human third molars and randomly divided into five groups (n = 10): (1) Control group treated with 37% phosphoric acid, (2) GD group, (3) Nd:YAG laser group (1064 nm, 100 µs, 10 Hz, 300 µm fiber, 1 W power, 100 mJ energy, and 85 J/cm2 energy density), (4) GD followed by Nd:YAG laser group, and (5) Nd:YAG laser followed by GD group. Scanning electron microscopy (SEM) was used to capture five images from each sample for analysis of dentinal tubules using Image J software. SEM/EDX elemental analysis was performed to determine the main mineral contents. Data analyzed using one-way ANOVA and Tukey's post hoc test for statistical comparisons. RESULTS: Laser and combination groups showed a significant decrease in dentinal tubule counts compared with the control and GD groups (p < 0.0001). There were no significant differences in open dentinal tubule counts between the control and GD groups, as well as between the laser and combination groups. However, significant differences were observed in the total area, average size of the tubules, and percentage area between the control group and the treatment groups (GD, laser, GD + laser, laser + GD). No significant difference was found in the Ca/P ratio between the tested groups. CONCLUSION: The use of Nd:YAG laser alone or in combination with GD was more effective in occluding dentinal tubules compared to GD alone. CLINICAL SIGNIFICANCE: This study has shown that Nd:YAG laser alone and in combination with GD has superior dentinal tubule occlusion in vitro. Its clinical use in the treatment of dentinal hypersensitivity may overcome the drawback of conventional treatment approaches for dentin hypersensitivity needing repeated applications to achieve continuous relief from pain since acidic diet and toothbrushing result in the continuing elimination of precipitates and surface coatings. How to cite this article: Alzarooni AH, El-Damanhoury HM, Aravind SS, et al. Combined Effects of Glutaraldehyde-based Desensitizer and Nd: YAG Laser on Dentinal Tubules Occlusion. J Contemp Dent Pract 2024;25(1):52-57.

Evaluation of Direct Antimicrobial Susceptibility Testing from Positive Flagged Blood Cultures in Sepsis Patients.

Background: Presently, many laboratories are equipped with automated system for antimicrobial susceptibility testing (AST) for minimum inhibitory concentration-based reporting which enables the clinician to choose the right antimicrobial for timely treatment of sepsis. The study aimed to assess performance of direct AST from blood culture positive broth using automated AST system for accuracy and time taken to release the report. Materials and methods: The present study conducted in a 25-bedded ICU in North India for 12 months. Single morphotype of bacteria on gram stain from positively flagged blood culture bottles were included, which was directly identified (using an in-house protocol) with MALDI-TOF-MS from positive blood culture broths. DAST was carried out from 200 such blood culture broths and results were compared with reference AST (RAST) which was also done using VITEK-2 using overnight grown bacterial colonies as per standard protocol. Results: Among 60 isolates of Enterobacterales, 99% categorical agreement for both E. coli and K. pneumoniae observed by two methods were tested for AST. Among non-fermenters, Pseudomonas aeruginosa showed a categorical agreement of 99.6%, as compared with Acinetobacter spp. and exotic GNBs, which showed 95-96% agreement. A significant difference of 18-24 hours was noted in time to release the report between DAST and RAST, for GNB and GPC both. Conclusion: Direct AST from positive flagged blood culture bottles can significantly reduce the time to release the bacterial susceptibility report by up to 24 hours, at the same time maintaining the accuracy. How to cite this article: Singh V, Agarwal J, Nath SS, Sharma A. Evaluation of Direct Antimicrobial Susceptibility Testing from Positive Flagged Blood Cultures in Sepsis Patients. Indian J Crit Care Med 2024;28(4):387-392.

Contradictory Recommendation in the Guideline for Antibiotic Prescription.

How to cite this article: Das PK, Nath SS, Parashar S. Contradictory Recommendation in the Guideline for Antibiotic Prescription. Indian J Crit Care Med 2024;28(7):713-714.

Unanswered Questions in the Guidelines for Antibiotic Prescription in Critically Ill Patients.

How to cite this article: Nath SS, Nachimuthu N, Bhagyashree, Singh S. Unanswered Questions in the Guidelines for Antibiotic Prescription in Critically Ill Patients. Indian J Crit Care Med 2024;28(7):715-716.

Are Deep Learning Structural Models Sufficiently Accurate for Virtual Screening? Application of Docking Algorithms to AlphaFold2 Predicted Structures.

Machine learning-based protein structure prediction algorithms, such as RosettaFold and AlphaFold2, have greatly impacted the structural biology field, arousing a fair amount of discussion around their potential role in drug discovery. While there are few preliminary studies addressing the usage of these models in virtual screening, none of them focus on the prospect of hit-finding in a real-world virtual screen with a model based on low prior structural information. In order to address this, we have developed an AlphaFold2 version where we exclude all structural templates with more than 30% sequence identity from the model-building process. In a previous study, we used those models in conjunction with state-of-the-art free energy perturbation methods and demonstrated that it is possible to obtain quantitatively accurate results. In this work, we focus on using these structures in rigid receptor-ligand docking studies. Our results indicate that using out-of-the-box Alphafold2 models is not an ideal scenario for virtual screening campaigns; in fact, we strongly recommend to include some post-processing modeling to drive the binding site into a more realistic holo model.

SABRE hyperpolarized anticancer agents for use in 1 H MRI.

PURPOSE: Enabling drug tracking (distribution/specific pathways) with magnetic resonance spectroscopy requires manipulation (via hyperpolarization) of spin state populations and targets with sufficiently long magnetic lifetimes to give the largest possible window of observation. Here, we demonstrate how the proton resonances of a group of thienopyridazines (with known anticancer properties), can be amplified using the para-hydrogen (p-H2 ) based signal amplification by reversible exchange (SABRE) hyperpolarization technique. METHODS: Thienopyridazine isomers, including a 2 H version, were synthesized in house. Iridium-based catalysts dissolved in a methanol-d4 solvent facilitated polarization transfer from p-H2 gas to the target thienopyridazines. Subsequent SABRE 1 H responses of hyperpolarized thienopyridazines were completed (400 MHz NMR). Pseudo-singlet state approaches were deployed to extend magnetic state lifetimes. Proof of principle spectral-spatial images were acquired across a range of field strengths (7T-9.4T MRI). RESULTS: 1 H-NMR signal enhancements of -10,130-fold at 9.4T (~33% polarization) were achieved on thieno[2,3-d]pyridazine (T[2,3-d]P), using SABRE under optimal mixing/field transfer conditions. 1 H T1 lifetimes for the thienopyridazines were ~18-50 s. Long-lived state approaches extended the magnetic lifetime of target proton sites in T[2,3-d]P from an average of 25-40 seconds. Enhanced in vitro imaging (spatial and chemical shift based) of target T[2,3-d]P was demonstrated. CONCLUSION: Here, we demonstrate the power of SABRE to deliver a fast and cost-effective route to hyperpolarization of important chemical motifs of anticancer agents. The SABRE approach outlined here lays the foundations for realizing continuous flow, hyperpolarized tracking of drug delivery/pathways.

Are Deep Learning Structural Models Sufficiently Accurate for Free-Energy Calculations? Application of FEP+ to AlphaFold2-Predicted Structures.

The availability of AlphaFold2 has led to great excitement in the scientific community─particularly among drug hunters─due to the ability of the algorithm to predict protein structures with high accuracy. However, beyond globally accurate protein structure prediction, it remains to be determined whether ligand binding sites are predicted with sufficient accuracy in these structures to be useful in supporting computationally driven drug discovery programs. We explored this question by performing free-energy perturbation (FEP) calculations on a set of well-studied protein-ligand complexes, where AlphaFold2 predictions were performed by removing all templates with >30% identity to the target protein from the training set. We observed that in most cases, the ΔΔG values for ligand transformations calculated with FEP, using these prospective AlphaFold2 structures, were comparable in accuracy to the corresponding calculations previously carried out using crystal structures. We conclude that under the right circumstances, AlphaFold2-modeled structures are accurate enough to be used by physics-based methods such as FEP in typical lead optimization stages of a drug discovery program.

Groundwater characterization and non-carcinogenic and carcinogenic health risk assessment of nitrate exposure in the Mahanadi River Basin of India.

Agriculture is the mainstay of India's economy and chemical fertilizers have been extensively used to meet increasing demands. Anthropogenic interventions at the soil surface, especially the application of nitrogenous fertilizers in agricultural fields, provide essential nutrients but become major pollutant sources in terrestrial ecosystems and aquatic environments. Groundwater samples from phreatic aquifers of the Mahanadi River Basin, Chhattisgarh, India, showed that the Ca2+-Mg2+-HCO3- freshwater type dominates, followed by the Ca2+-Mg2+-Cl- and Na+-HCO3- types. Increasing trends in the ionic ratios of (NO3-+Cl-)/HCO3- over TDS and of NO3-/Cl- over Cl- indicated the significant impact of anthropogenic pollution on groundwater contamination. Deterministic and probabilistic approaches were used to assess the non-carcinogenic and carcinogenic health risks of nitrate to children and adults. Both approaches produced the same results and indicated children were more prone to non-carcinogenic health risk than adults. An excess gastric cancer risk (ER) exposure model showed that approximately 42% of the groundwater samples had a non-negligible ER (1.00 × 10-4 to 1.00 × 10-5). Sensitivity analysis indicated groundwater nitrate concentration, ingestion rate, and the percentage of nitrite from nitrate were the most significant variables in determining HI and ER. It is suggested to adopt proper management of control policies for reducing the elevated groundwater nitrate concentration in the present study area.

Targeted Degradation of the Oncogenic Phosphatase SHP2.

SHP2 is a protein tyrosine phosphatase that plays a critical role in the full activation of the Ras-MAPK pathway upon stimulation of receptor tyrosine kinases, which are frequently amplified or mutationally activated in human cancer. In addition, activating mutations in SHP2 result in developmental disorders and hematologic malignancies. Several allosteric inhibitors have been developed for SHP2 and are currently in clinical trials. Here, we report the development and evaluation of a SHP2 PROTAC created by conjugating RMC-4550 with pomalidomide using a PEG linker. This molecule is highly selective for SHP2, induces degradation of SHP2 in leukemic cells at submicromolar concentrations, inhibits MAPK signaling, and suppresses cancer cell growth. SHP2 PROTACs serve as an alternative strategy for targeting ERK-dependent cancers and are useful tools alongside allosteric inhibitors for dissecting the mechanisms by which SHP2 exerts its oncogenic activity.

Ror ß expression in activated macrophages and dental pulp stem cells.

AIM: To assess the expression of Retinoic acid-related orphan receptor beta (Ror ß) in human inflamed dental pulp stem cells (hI-DPSCs) and during macrophage phenotypic conversion. METHODOLOGY: Commercially procured THP-1 monocytes conversion to macrophages was judged by their morphology, the percentage of adherent cells and the expression of CD-14 surface marker. THP-1 macrophage cell viability following LPS, IFN-γ/IL-4, IL-13 stimulus was evaluated at 24 and 48h. The phenotypic conversion of macrophages to M1 and M2 was confirmed by flow cytometry and Western blot analysis. Cytokine release following polarization was estimated by the BD cytokine flex kit. The expression of Ror ß in THP-1 macrophages and hI-DPSCs following LPS, IFN-γ/IL-4, IL-13 stimulus was assessed by Western blot analysis. Statistical significance was analysed using one-way Anova followed by Tukey's Post hoc test. RESULTS: THP-1 monocytes pretreated with PMA (100 ng mL-1 ) for 48 h followed by culturing in PMA-free media for another 48 h yielded cells with morphological characteristics similar to macrophages with a high percentage of adherence capability and CD-14 expression. Macrophages treated with LPS 100 ng mL-1 and IFN-γ 20 ng mL-1 or IL-4 20 ng mL-1 had high expression of the respective M1 and M2 CD markers in flow cytometry and Western blot analysis. Cytokine release studies demonstrated the expression of IL-1ß, TNF-α and IL-10 in the M1-polarized macrophages (P < 0.01), whilst TGF- ß levels were seen in the M1 and M2-polarized macrophages. Ror ß expression was upregulated when macrophages and hI-DPSCs were treated with anti-inflammatory cytokines. CONCLUSION: Ror ß was expressed in THP-1 macrophages and hI-DPSCs during their resting stage. Upregulated expression of Ror ß occurred following an anti-inflammatory stimulus.

15 N hyperpolarisation of the antiprotozoal drug ornidazole by Signal Amplification By Reversible Exchange in aqueous medium.

Signal amplification by reversible exchange (SABRE) offers a cost-effective route to boost nuclear magnetic resonance (NMR) signal by several orders of magnitude by employing readily available para-hydrogen as a source of hyperpolarisation. Although 1 H spins have been the natural choice of SABRE hyperpolarisation since its inception due to its simplicity and accessibility, limited spin lifetimes of 1 H makes it harder to employ them in a range of time-dependent NMR experiments. Heteronuclear spins, for example, 13 C and 15 N, in general have much longer T1 lifetimes and thereby are found to be more suitable for hyperpolarised biological applications as demonstrated previously by para-hydrogen induced polarisation (PHIP) and dynamic nuclear polarisation (DNP). In this study we demonstrate a simple procedure to enhance 15 N signal of an antibiotic drug ornidazole by up to 71,000-folds with net 15 N polarisation reaching ~23%. Further, the effect of co-ligand strategy is studied in conjunction with the optimum field transfer protocols and consequently achieving 15 N hyperpolarised spin lifetime of >3 min at low field. Finally, we present a convenient route to harness the hyperpolarised solution in aqueous medium free from catalyst contamination leading to a strong 15 N signal detection for an extended duration of time.

Comparison of the Efficacy of Different Arterial Waveform-derived Variables (Pulse Pressure Variation, Stroke Volume Variation, Systolic Pressure Variation) for Fluid Responsiveness in Hemodynamically Unstable Mechanically Ventilated Critically Ill Patients.

INTRODUCTION: This study was conducted to assess fluid responsiveness in critically ill patients to avoid various complications of fluid overload. MATERIAL AND METHODS: This study was done in an ICU of a tertiary care hospital after approval from the institute ethical committee over 18 months. A total of 54 consenting adult patients were included in the study. Patients were hemodynamically unstable requiring mechanical ventilation, had acute circulatory failure, or those with at least one clinical sign of inadequate tissue perfusion. All patients were ventilated using tidal volume of 6-8 mL/kg, RR-12-15/minutes, positive end expiratory pressure (PEEP)-5 cm of water, and plateau pressure was kept below 30 cm water. They were sedated throughout the study. The arterial line and the central venous catheter were placed and connected to Vigileo-FloTrac transducer (Edward Lifesciences). Patients were classified into responder and nonresponder groups on the basis of the cardiac index (CI) after fluid challenge of 10 mL/kg of normal saline over 30 minutes. Pulse pressure variation (PPV), stroke volume variation (SVV), and systolic pressure variation (SPV) were assessed and compared at baseline, 30 minutes, and 60 minutes. RESULTS: In our study we found that PPV and SVV were significantly lower among responders than nonresponders at 30 minutes and insignificant at 60 minutes. Stroke volume variation was 10.28 ± 1.76 in the responder compared to 12.28 ± 4.42 (p = 0.02) at 30 minutes and PPV was 15.28 ± 6.94 in responders while it was 20.03 ± 4.35 in nonresponders (p = 0.01). We found SPV was insignificant at all time periods among both groups. CONCLUSION: We can conclude that initial assessment for fluid responsiveness in critically ill mechanically ventilated patients should be based on PPV and SVV to prevent complications of fluid overload and their consequences. HOW TO CITE THIS ARTICLE: Kumar N, Malviya D, Nath SS, Rastogi S, Upadhyay V. Comparison of the Efficacy of Different Arterial Waveform-derived Variables (Pulse Pressure Variation, Stroke Volume Variation, Systolic Pressure Variation) for Fluid Responsiveness in Hemodynamically Unstable Mechanically Ventilated Critically Ill Patients. Indian J Crit Care Med 2021;25(1):48-53.

Reversing the Trend of Antimicrobial Resistance in ICU: Role of Antimicrobial and Diagnostic Stewardship.

BACKGROUND: Increasing antimicrobial resistance (AMR) among common bacteria combined with the slow development of new antibiotics has posed a challenge to clinicians. AIM AND OBJECTIVE: To demonstrate whether antimicrobial and diagnostic stewardship program (ASP and DSP)-related interventions improve antibiotic susceptibilities among common bacteria causing bloodstream infections (BSI) in patients admitted to the intensive care unit (ICU) and whether these resulted in changes in the volume of antimicrobial consumption. MATERIALS AND METHODS: We compared the susceptibility patterns of gram-negative bacteria (GNB) and gram-positive cocci (GPC) causing BSI and changes in the volume of antibiotics prescribed for the same before and after 2017 by a retrospective analysis. RESULTS: Postintervention, there was increased susceptibility of all GNBs to aminoglycosides; Escherichia coli and Klebsiella spp. to beta-lactambeta-lactamase inhibitors (BLBLI) combinations; and Klebsiella spp. and Pseudomonas spp. to carbapenems. Acinetobacter spp., Klebsiella spp., and Pseudomonas spp. showed improved susceptibility to doxycycline, whereas E. coli and Klebsiella spp. showed significantly improved susceptibility to fluoroquinolones. Among GPCs, there was increased susceptibility of Staphylococcus aureus (levofloxacin, clindamycin, and aminoglycoside), coagulase-negative S. aureus (CoNS) (chloramphenicol, levofloxacin, clindamycin, and aminoglycoside), and enterococci (chloramphenicol, levofloxacin, and clindamycin). There was a significant reduction in usage of antimicrobials for the treatment of GPCs (linezolid, doxycycline, chloramphenicol, levofloxacin, BLBLI, macrolide, and cephalosporin) and GNBs (levofloxacin, cephalosporin, carbapenem, and colistin), which caused BSI. CONCLUSION: The present study illustrated that combined ASP and DSP interventions successfully reversed the resistance pattern of organisms causing BSI and resulted in a reduction in antibiotic utilization. HOW TO CITE THIS ARTICLE: Agarwal J, Singh V, Das A, Nath SS, Kumar R, Sen M. Reversing the Trend of Antimicrobial Resistance in ICU: Role of Antimicrobial and Diagnostic Stewardship. Indian J Crit Care Med 2021;25(6):635-641.

A comparative study evaluating the dose volume parameters in 3D conformal radiation of left sided whole breast irradiation including regional lymphnodes - a need of resource constrained countries.

BACKGROUND: The purpose of this study was to compare four 3D conformal radiation techniques in treatment of left breast cancer patients. MATERIALS AND METHODS: Radiation was planned for 20 patients to the left breast and regional lymph nodes using four techniques: partially wide tangents, photon-photon mix, photon-electron mix and 30/70 photon-electron mix. All plans were evaluated for internal mammary nodes (IMN) coverage, hotspot and normal tissue constraints. RESULT: The 85% of planning target volume (PTV) coverage was lesser for upper IMN than the lower IMN (below the lower border of the clavicular head) for all four techniques. The lower IMN coverage was better for partially wide tangent (80.46%) and photon-photon mix (88.88%). The lowest value of hotspot was seen in the partially wide tangent technique (112.69% ± 1.92). Hotspot is unacceptably high in both photon-electron mix and 30/70 photon-electron mix (> 120%). Left lung mean dose for all techniques on a pair-wise comparison showed no statistical difference. Left lung V20 values for partially wide tangent was 37.56% ± 8.17 and for photon-photon mix it was 40.49% ± 3.36. The mean heart dose with partially wide tangent was 9.43 ± 3.15 Gy and with photon-photon mix it was 10.10 ± 2.70 Gy. The mean heart dose for photon-electron mix was 7.56 ± 1.95 Gy and for 30/70 photon-electron mix it was 7.98 ± 2.16 Gy. CONCLUSION: No single technique satisfies all the criteria. The decision should be made on a case-by-case basis, considering the anatomy of the patient, availability of electron facilities and setup accuracy and reproducibility.

A role for low concentration reaction intermediates in the signal amplification by reversible exchange process revealed by theory and experiment.

A route to monitor the involvement of less abundant species during the catalytic transfer of hyperpolarisation from parahydrogen into a substrate is detailed. It involves probing how the degree of hyperpolarisation transfer catalysis is affected by the magnetic field experienced by the catalyst during this process as a function of temperature. The resulting data allow the ready differentiation of the roles played by hard to detect and highly reactive complexes, such as [Ir(H)2(NHC)(substrate)2(methanol)]Cl, from dominant species such as [Ir(H)2(NHC)(substrate)3]Cl. The difference in behaviour results from changes in the interligand spin-spin coupling network within the active SABRE catalysts.

A simple and cost-efficient technique to generate hyperpolarized long-lived 15N-15N nuclear spin order in a diazine by signal amplification by reversible exchange.

Signal Amplification by Reversible Exchange (SABRE) is an inexpensive and simple hyperpolarization technique that is capable of boosting nuclear magnetic resonance sensitivity by several orders of magnitude. It utilizes the reversible binding of para-hydrogen, as hydride ligands, and a substrate of interest to a metal catalyst to allow for polarization transfer from para-hydrogen into substrate nuclear spins. While the resulting nuclear spin populations can be dramatically larger than those normally created, their lifetime sets a strict upper limit on the experimental timeframe. Consequently, short nuclear spin lifetimes are a challenge for hyperpolarized metabolic imaging. In this report, we demonstrate how both hyperpolarization and long nuclear spin lifetime can be simultaneously achieved in nitrogen-15 containing derivatives of pyridazine and phthalazine by SABRE. These substrates were chosen to reflect two distinct classes of 15N2-coupled species that differ according to their chemical symmetry and thereby achieve different nuclear spin lifetimes. The pyridazine derivative proves to exhibit a signal lifetime of ∼2.5 min and can be produced with a signal enhancement of ∼2700. In contrast, while the phthalazine derivative yields a superior 15 000-fold 15N signal enhancement at 11.7 T, it has a much shorter signal lifetime.

Osteogenic and odontogenic differentiation potential of dental pulp stem cells isolated from inflamed dental pulp tissues (I-DPSCs) by two different methods.

Objective: The objective of the present study is to isolate stem cells from inflamed dental pulp tissues (I-DPSCs) and study the characteristic such as surface markers, osteo/odontogenic differentiation potential between the outgrowth (OG) and enzymatic digestion (COL) methods.Materials and methods: I-DPSCs harvested by both methods were analysed for Mesenchymal Stem Cell marker expression by flow cytometry. The metabolic activity of the isolated cells was assessed by MTT assay. The Alkaline Phosphatase (ALP) and Alizarin red staining was done to analyse the osteogenic potential of isolated cells. The osteo/odontogenic differentiation was done by checking the expression of Dentine Matrix Protein 1 (DMP1), Dentine Sialophosphoprotein (DSPP), ALP and Bone Gamma-Carboxyglutamate Protein (BGLAP) by Real time PCR.Results: The isolated cells were positive for MSC markers such as CD-90, CD-105 and CD-73 and negative for CD-14, CD-45 and STRO-1. MTT assay indicated that the I-DPSCs from OG method showed higher metabolic activity than cells from COL. However, the osteo/odontogenic differentiation was in favour of cells isolated by COL method.Conclusion: Although the cell metabolic rate was more in OG, the osteo/odontogenic differentiation was higher in COL, suggesting that the isolation method and culture conditions do affect the differentiation capacity of isolated cells.

Iridium α-Carboxyimine Complexes Hyperpolarized with para-Hydrogen Exist in Nuclear Singlet States before Conversion into Iridium Carbonates.

The formation and hyperpolarization of an [Ir(H)2 (amine)(IMes)(η2 -imine)]Cl complex that can be created in a hyperpolarized nuclear singlet state is reported. These complexes are formed when an equilibrium mixture of pyruvate, amine (benzylamine or phenylethylamine), and the corresponding imine condensation product, react with preformed [Ir(H)2 (amine)3 (IMes)]Cl. These iridium α-carboxyimine complexes exist as two regioisomers differentiated by the position of amine. When examined with para-hydrogen the hydride resonances of the isomer with amine trans to hydride become strongly hyperpolarized. The initial hydride singlet states readily transfer to the corresponding 13 C2 state in the labelled imine and exhibit magnetic state lifetimes of up to 11 seconds. Their 13 C signals have been detected with up to 420 fold signal gains at 9.4 T. On a longer timescale, and in the absence of H2 , further reaction leads to the formation of neutral carbonate containing [Ir(amine)(η2 -CO3 )(IMes)(η2 -imine)]. Complexes are characterized by, IR, MS, NMR and X-ray diffraction.

Halitosis and Pain Post Electrocautery Adenoidectomy.

Background and Objectives: Electrocautery adenoidectomy (ECA) is a common procedure performed in paediatric otolaryngology. ECA has been preferred over curettage adenoidectomy due to its lower intraoperative bleeding rates, decreased procedure time, and higher subjective success. However, post-ECA symptoms of pain and halitosis have never been studied. The objective of our study was to identify the pattern of post-ECA halitosis and pain in the paediatric population. Materials and Methods: This is a single centre, prospective observational study that uses visual analogue scales (VAS) by parent proxy to assess post-ECA pain and halitosis in paediatric patients (age < 18) in South Australia. A total of 19 patients were enrolled in the study and followed for seven days. Results: Postoperative pain and halitosis reaches a peak 3 days post-ECA (median = 2 for pain; median = 6 for halitosis) but resolves 7 days post-ECA (median = 0 for both). Conclusions: Our study demonstrates that halitosis and pain occur over a seven-day period in patients undergoing ECA and will resolve post-operatively with simple analgesia and without antibiotics.