Phytocompounds screening of Nigella sativa in terms of human cancer by targeting sphingosine kinase-1 and pyruvate kinase-M2: a study based on in silico analysis.

Cancer is a multifactorial disease that can cause morbidity and mortality in humans. An altered gene expression in cancer leads to a change in the overall activity of the human cell. Overexpression of cancer protein may give a piece of wide information about the specific type of tumor. Sphingosine kinase-1 (SK-1) is a metabolic enzyme that is mainly overexpressed in several types of cancer and other inflammatory diseases. Similarly, pyruvate kinase-M2 (PK-M2) is an important oncogenic ATP-producing glycolytic enzyme that is upregulated in most cancer cells. The phytocompound of medicinal plants such as Nigella sativa contains a variety of micronutrients that inhibit the proliferation and activity of tumor cells. In this study, the role of phytocompounds in combating cancer was studied against the model kinase proteins, that is, PK-M2 and SK-1. In silico tool like the PASS-Way2Drug server was used to predict the anticancer properties of phytocompounds. Moreover, the CLC-Pred web server provided the cytotoxicity prediction of chemical compounds against several human cancer cell lines. The pharmacokinetics and toxicity profiles were predicted by the SwissADME and pkCSM software. The binding energies were obtained by molecular docking to confirm the intermolecular interaction of selected phytocompounds with proteins. Consequently, molecular dynamics (MD) simulation confirmed the stability, conformational changes, and dynamic behavior of the kinase proteins complexed with the lead phytocompounds, that is, epicatechin, apigenin, and kaempferol.Communicated by Ramaswamy H. Sarma.

Preparation of nanoformulation of 5-fluorouracil to improve anticancer efficacy: integrated spectroscopic, docking, and MD simulation approaches.

Nanoformulations (NFs) can be used as a novel drug delivery system to treat all cancer types. One of the major drawbacks of conventional anticancer drugs is that they have poor specificity and higher toxicity towards normal cells. 5-fluorouracil (5-FU) is a well-studied anticancer drug that has a significant role in various cancers, specifically colorectal cancer therapy. This study was performed to determine the functional groups, particle size, surface charge, heterogeneity, and stability of the NF. The NFs of 5-FU were prepared through the ultrasonication technique by increasing the surfactant (Tween-80) concentrations. Among all three NFs, nanoformulated 5-FU (n5-FU) showed the most effective particle size (10.72 nm) with a zeta potential of (-4.57 mV). The cytotoxicity and apoptosis profiles confirmed that n5-FU enhanced the anticancer effect of the pure drug in HCT-116 cells, as evident from MTT assay, fluorescence microscopy, and FACS analysis. In HCT-116 cells, the IC50 values of pure and n5-FU were obtained as 41.3 µM and 18.8 µM, respectively, indicating that n5-FU was more effective against the cancer cell line. The cellular uptake study was performed to check the intake of NF in cancer cells. However, the microtubule-affinity regulating kinase-4 (MARK-4), a cancer-target protein, was purified to study the inhibition and interaction studies. The inhibition assay confirmed the inhibitory potential of 5-FU against MARK-4 protein. the multi-spectroscopic, molecular docking and MD simulation studies were performed to analyse the conformational changes, binding studies, intermolecular interactions, and stability of MARK-4 protein upon binding 5-FU. This demonstrates that NF can enhance the effectiveness of anticancer drugs.Communicated by Ramaswamy H. Sarma.

A placebo-controlled randomized clinical trial of antibiotics versus probiotics as an adjuvant to nonsurgical periodontal treatment among smokers with Stage III, Grade C generalized periodontitis.

BACKGROUND: Probiotics are viable microorganisms, which if delivered in appropriate dose can provide health benefits. Lactobacillus reuteri (DM17938+ATCC PTA 5289) has been recommended as a safe choice for probiotics. The objective of this study is to compare the improvement in the periodontal parameters amongst smokers with generalized periodontitis with Stage III, Grade C treated with nonsurgical periodontal treatment (NSPT) to which either an antibiotics or probiotics were given as an adjuvant. METHODS: Sixty smokers with Stage III, Grade C generalized periodontitis were randomized in two groups after taking informed consent. Periodontal parameters including bleeding on probing (BOP), probing depth (PD), attachment loss (AL), gingival index (GI), and plaque index (PI) were recorded. Group 1 received (after NSPT and oral hygiene instructions) amoxicillin and metronidazole for 7 days and a placebo for probiotics for 30 days. Group 2 was provided (after NSPT and oral hygiene instructions) with one tablet of Lactobacillus reuteri probiotics (2 × 108 CFU) twice daily for 30 days and placebo antibiotics for 7 days. The periodontal parameters were recorded again at 1- and 3-month follow-ups as outcome variables. Mean, standard deviation, and confidence interval were reported using SPSS 20.0. RESULTS: A statistically significant clinical improvement in the PD, BOP, PI, and GI were observed in both the groups at 3-month follow-up. However, the AL remained unchanged in both the groups. CONCLUSIONS: Administration of probiotics and antibiotics along with NSPT yield statistically significant differences in PD and BOP from baseline to 3-month follow-up. However, between the group differences were not statistically significant for the periodontal parameters (AL, PD, and BOP).

Structural and functional insights into Spns2-mediated transport of sphingosine-1-phosphate.

Sphingosine-1-phosphate (S1P) is an important signaling sphingolipid that regulates the immune system, angiogenesis, auditory function, and epithelial and endothelial barrier integrity. Spinster homolog 2 (Spns2) is an S1P transporter that exports S1P to initiate lipid signaling cascades. Modulating Spns2 activity can be beneficial in treatments of cancer, inflammation, and immune diseases. However, the transport mechanism of Spns2 and its inhibition remain unclear. Here, we present six cryo-EM structures of human Spns2 in lipid nanodiscs, including two functionally relevant intermediate conformations that link the inward- and outward-facing states, to reveal the structural basis of the S1P transport cycle. Functional analyses suggest that Spns2 exports S1P via facilitated diffusion, a mechanism distinct from other MFS lipid transporters. Finally, we show that the Spns2 inhibitor 16d attenuates the transport activity by locking Spns2 in the inward-facing state. Our work sheds light on Spns2-mediated S1P transport and aids the development of advanced Spns2 inhibitors.

A technical note on percutaneous fixation of trapezium fracture under gravitational traction.

Trapezium fractures rarely present as isolated fractures and warrant anatomical fixation to minimize post-traumatic arthritis. Fixation techniques reported include open approach to trapezium fracture and fixation, percutaneous fixation under image intensifier guidance or percutaneous arthroscopic fixation under gravitational traction. We have reported a novel method of percutaneous fixation of a trapezium Walker type IV coronal split fractures under gravitational traction and per-operative image intensifier guidance. Two percutaneous headless differential pitch screws were used for fixation, which was off-loaded using an inter-metacarpal K-wire. Patient was discharged on the same day with a light-weight cast. The fixation yielded acceptable reduction of the articular surface as confirmed by radiographs. The cast and off-loading K-wire were removed at four weeks post-surgery and hand therapy was commenced. 3-month review showed optimal radiological healing with excellent return to function without pain. In conclusion, using gravitational traction can allow percutaneous fixation of trapezium fractures with excellent results and also make an otherwise complex procedure simpler. LEVEL OF EVIDENCE: IV.

Nizatidine interacts with ct-DNA causing genotoxicity and cytotoxicity: an assessment by in vitro, in vivo, and in silico studies.

H2 receptor antagonists are the medication given for treating stomach ulcers, but lately, reports have shown their role in healing several malignant ulcers. The present work entails the interaction of H2 blocker nizatidine with calf thymus (ct)-DNA for determining the binding mode and energetics of the interaction. Multi-spectroscopic, calorimetric, viscometric and bioinformatic analysis revealed that nizatidine interacted with ct-DNA via groove-binding mode and is characterised by exothermic reaction. Moreover, assessment of genotoxic potential of nizatidine in vitro was carried out in peripheral human lymphocytes by alkaline comet assay. DNA damage occurred at high concentrations of nizatidine. Genotoxicity of nizatidine was also evaluated in vivo by assessing cytogenetic biomarkers viz. micronuclei formation and chromosomal aberration test. Nizatidine was able to induce micronuclei formation and chromosomal damage at high dose. Additionally, cytotoxic activity of nizatidine was determined in cancer cell lines, namely HeLa and HCT-116 and compared with the normal human cell line HEK-293 employing MTT assay. It was observed that nizatidine was more toxic towards HeLa and HCT-116 than HEK-293. Cell morphology analysis by compound inverted microscopy further strengthens the finding obtained through MTT assay.

Combining cysteine scanning with chemical labeling to map protein-protein interactions and infer bound structure in an intrinsically disordered region.

The Mycobacterium tuberculosis genome harbours nine toxin-antitoxin (TA) systems of the mazEF family. These consist of two proteins, a toxin and an antitoxin, encoded in an operon. While the toxin has a conserved fold, the antitoxins are structurally diverse and the toxin binding region is typically intrinsically disordered before binding. We describe high throughput methodology for accurate mapping of interfacial residues and apply it to three MazEF complexes. The method involves screening one partner protein against a panel of chemically masked single cysteine mutants of its interacting partner, displayed on the surface of yeast cells. Such libraries have much lower diversity than those generated by saturation mutagenesis, simplifying library generation and data analysis. Further, because of the steric bulk of the masking reagent, labeling of virtually all exposed epitope residues should result in loss of binding, and buried residues are inaccessible to the labeling reagent. The binding residues are deciphered by probing the loss of binding to the labeled cognate partner by flow cytometry. Using this methodology, we have identified the interfacial residues for MazEF3, MazEF6 and MazEF9 TA systems of M. tuberculosis. In the case of MazEF9, where a crystal structure was available, there was excellent agreement between our predictions and the crystal structure, superior to those with AlphaFold2. We also report detailed biophysical characterization of the MazEF3 and MazEF9 TA systems and measured the relative affinities between cognate and non-cognate toxin-antitoxin partners in order to probe possible cross-talk between these systems.

A bi-institutional multi-disciplinary failure mode and effects analysis (FMEA) for a Co-60 based total body irradiation technique.

BACKGROUND: We aim to assess the risks associated with total body irradiation (TBI) delivered using a commercial dedicated Co-60 irradiator, and to evaluate inter-institutional and inter-professional variations in the estimation of these risks. METHODS: A failure mode and effects analysis (FMEA) was generated using guidance from the AAPM TG-100 report for quantitative estimation of prospective risk metrics. Thirteen radiation oncology professionals from two institutions rated possible failure modes (FMs) for occurrence (O), severity (S), and detectability (D) indices to generate a risk priority number (RPN). The FMs were ranked by descending RPN value. Absolute gross differences (AGD) in resulting RPN values and Jaccard Index (JI; for the top 20 FMs) were calculated. The results were compared between professions and institutions. RESULTS: A total of 87 potential FMs (57, 15, 10, 3, and 2 for treatment, quality assurance, planning, simulation, and logistics respectively) were identified and ranked, with individual RPN ranging between 1-420 and mean RPN values ranging between 6 and 74. The two institutions shared 6 of their respective top 20 FMs. For various institutional and professional comparison pairs, the number of common FMs in the top 20 FMs ranged from 6 to 13, with JI values of 18-48%. For the top 20 FMs, the trend in inter-professional variability was institution-specific. The mean AGD values ranged between 12.5 and 74.5 for various comparison pairs. AGD values differed the most for medical physicists (MPs) in comparison to other specialties i.e. radiation oncologists (ROs) and radiation therapists (RTs) [MPs-vs-ROs: 36.3 (standard deviation SD = 34.1); MPs-vs-RTs: 41.2 (SD = 37.9); ROs-vs-RTs: 12.5 (SD = 10.8)]. Trends in inter-professional AGD values were similar for both institutions. CONCLUSION: This inter-institutional comparison provides prospective risk analysis for a new treatment delivery unit and illustrates the institution-specific nature of FM prioritization, primarily due to operational differences. Despite being subjective in nature, the FMEA is a valuable tool to ensure the identification of the most significant risks, particularly when implementing a novel treatment modality. The creation of a bi-institutional, multidisciplinary FMEA for this unique TBI technique has not only helped identify potential risks but also served as an opportunity to evaluate clinical and safety practices from the perspective of both multiple professional roles and different institutions.

Monte Carlo commissioning of radiotherapy LINAC-Introducing an improved methodology.

PURPOSE: Monte Carlo (MC) commissioning of medical linear accelerator (LINAC) is a time-consuming process involving a comparison between measured and simulated cross beam/lateral profiles and percentage depth doses (PDDs) for various field sizes. An agreement between these two data sets is sought by trial and error method while varying the incident electron beam parameters, such as electron beam energy or width, etc. This study aims to improve the efficiency of MC commissioning of a LINAC by assessing the feasibility of using a limited number of simulated PDDs. MATERIALS AND METHODS: Using EGSnrc codes, a Varian Clinac 2100 unit has been commissioned for 6 MV photon beam, and a methodology has been proposed to identify the incident electron beam parameters in a speedier fashion. Impact of voxel size in 3-dimensions and cost functions used for comparison of the measured and simulated data have been investigated along with the role of interpolation. RESULTS: A voxel size of 1 × 1×0.5 cm3 has been identified as suitable for accurate and fast commissioning of the LIANC. The optimum number of simulated PDDs (required for further interpolation) has been found to be five. CONCLUSION: The present study suggests that PDDs alone at times can be insufficient for an unambiguous commissioning process and should be supported by including the lateral beam profiles in the process.

Arthroscopic knotless anterior labral stabilization using labral tape and wide awake anaesthesia-short term results.

BACKGROUND: The shoulder is the least constrained of all joints of the body and is more susceptible to injury including dislocation. The rate of recurrent instability following primary stabilization procedure at 10 years of follow-up ranged from 3.4 to 20%. There is a lack of evidence in the literature regarding use of labral tape and anchors for anterior stabilization despite the growing market for this product. We describe the outcomes of 67 patients who underwent knotless arthroscopic anterior stabilisation under awake anaesthesia using 1.5 mm LabralTape with 2.9 mm Pushlock anchors for primary anterior instability by a single surgeon. METHODS: This was a retrospective analysis of prospectively collected outcome data for adult patients undergoing anterior stabilisation for primary traumatic anterior shoulder instability between 2013 and 2016 at two centres. Patients with > 25% glenoid bone loss, engaging Hill Sach's, and multidirectional instability were excluded. All cases underwent surgery using awake anaesthetic technique. The surgical technique and post-operative physiotherapy was standardized. Outcomes were measured at 6 months and 12 months. RESULTS: Of the 74 patients in our study, 7 were lost to follow up. Outcomes were measured using the Oxford Instability Shoulder Score (OISS) and clinical assessment including the range of motion. The OISS showed statistically significant improvement from a mean score and standard deviation (SD) of 24.72 ± 2.8 pre-surgery to 43.09 ± 3.5 after the procedure at 12 months with good to excellent outcomes in 66 cases (98.5%). The mean abduction was 134.2 ± 6.32 and external rotation was 72.55 ± 5.42 at 60-90 position at 12 months. We report no failures due to knot slippage or anchor pull-out. CONCLUSION: Our case series using the above technique has distinct advantages of combining a small non-absorbable implant with flat, braided, and high-strength polyethylene tape. This technique demonstrates superior medium term results to conventional suture knot techniques for labral stabilization thereby validating its use.

Translating bed total body irradiation lung shielding and dose optimization using asymmetric MLC apertures.

A revised translating bed total body irradiation (TBI) technique is developed for shielding organs at risk (lungs) to tolerance dose limits, and optimizing dose distribution in three dimensions (3D) using an asymmetrically-adjusted, dynamic multileaf collimator. We present a dosimetric comparison of this technique with a previously developed symmetric MLC-based TBI technique. An anthropomor-phic RANDO phantom is CT scanned with 3 mm slice thickness. Radiological depths (RD) are calculated on individual CT slices along the divergent ray lines. Asymmetric MLC apertures are defined every 9 mm over the phantom length in the craniocaudal direction. Individual asymmetric MLC leaf positions are optimized based on RD values of all slices for uniform dose distributions. Dose calculations are performed in the Eclipse treatment planning system over these optimized MLC apertures. Dose uniformity along midline of the RANDO phantom is within the confidence limit (CL) of 2.1% (with a confidence probability p = 0.065). The issue of over- and underdose at the interfaces that is observed when symmetric MLC apertures are used is reduced from more than ± 4% to less than ± 1.5% with asymmetric MLC apertures. Lungs are shielded by 20%, 30%, and 40% of the prescribed dose by adjusting the MLC apertures. Dose-volume histogram analysis confirms that the revised technique provides effective lung shielding, as well as a homogeneous dose coverage to the whole body. The asymmetric technique also reduces hot and cold spots at lung-tissue interfaces compared to previous symmetric MLC-based TBI technique. MLC-based shielding of OARs eliminates the need to fabricate and setup cumbersome patient-specific physical blocks.

Micromechanical Intervention in Sandwich Restoration.

OBJECTIVE: To evaluate the sealing ability of two different types of Glass Ionomer Cements (GICs) used for sandwich restorations and assess the effect of acid etching of GIC on microleakage at GIC-resin composite interface. STUDY DESIGN: Experimental study. PLACE AND DURATION OF STUDY: Department of Operative, DIEKIOHS (DUHS) and NED University, Karachi, from February to June 2011. METHODOLOGY: Eighty cavities were prepared on the proximal surfaces of 40 permanent human premolars (2 cavities per tooth), assigned to 4 groups (n=20) and restored as follows: Group CIE - Conventional GIC (CI) was applied onto the axial and cervical cavity walls, allowed setting for 5 minutes and acid etched (E) along the cavity margins with 37% phosphoric acid for 15 seconds, washed for 30 seconds and dried; the adhesive system was applied and light cured for 10 seconds, completing the restoration with composite resin light cured for 40 seconds; Group CIN - same as Group CIE, except for acid etching of the CI surface; Group RME - same as CIE, but using a resin modified GIC (RMGIC); Group RMN - same as Group RME, except for acid etching of the RMGIC surface. Specimens were soaked in 1% methylene blue dye solution at 37ºC for 24 hours, rinsed under running water for 15 minutes, bisected mesiodistally and dye penetration was measured following the ISO/TS 11405-2003 standard. Kruskal Wallis and post Hoc tests significant differences in the microleakage among all the four groups. RESULTS: There was a significant difference between the two groups of GICs (RMGIC and CI, p=0.001). There was no significant difference in between the two sub-groups that is between CIN and CIE (p=0.656), and between Groups RME and RMN (p=0.995). CONCLUSION: Phosphoric acid etching of GIC, prior to the placement of composite resin, does not improve the sealing ability of sandwich restorations. RMGIC was more effective in preventing dye penetration at the GIC-resin composite dentine interfaces than CI.

Validation of the femoral anteversion measurement method used in imageless navigation.

Total hip arthroplasty restores lost mobility to patients suffering from osteoarthritis and acute trauma. In recent years, navigated surgery has been used to control prosthetic component placement. Furthermore, there has been increasing research on what constitutes correct placement. This has resulted in the definition of a safe-zone for acetabular cup orientation. However, there is less definition with regard to femoral anteversion and how it should be measured. This study assesses the validity of the femoral anteversion measurement method used in imageless navigation, with particular attention to how the neutral rotation of the femur is defined. CT and gait analysis methodologies are used to validate the reference which defines this neutral rotation, i.e., the ankle epicondyle piriformis (AEP) plane. The findings of this study indicate that the posterior condylar axis is a reliable reference for defining the neutral rotation of the femur. In imageless navigation, when these landmarks are not accessible, the AEP plane provides a useful surrogate to the condylar axis, providing a reliable baseline for femoral anteversion measurement.

Fall-related fractures in persons with spinal cord impairment: a descriptive analysis.

Falls are a significant cause of injury, disability, and death in the elderly, but little is known about the risk of wheelchair-related falls. The purpose of this study is to describe the incidence, etiology, location of fracture, treatment, and health care utilization of fall-related fractures in persons with spinal cord impairment (SCI). A retrospective review of 45 medical records of patients with SCI who sustained fractures, nonconcomitant with the onset of their initial injury, was completed at a Veterans Health Administration (VHA) SCI service over a 10-year period. Of the 24 veterans who sustained fall-related fractures, three (12%) were found to have repeated falls with fractures. Falls were sustained during activities (more than one wheelchair activity contributed to a fall; e.g., transfer activity with brake failure in a van) including transfer (44%), reaching (11%), propelling (15%), moving in bed (22%), transferring or riding in a vehicle (30%), and showering (7%). Factors contributing to falls included loss of balance, equipment failure, muscle spasms, excessive speed, not wearing protective straps, and narcolepsy. Among the 31 fractures sustained in 27 fall episodes in 24 subjects, lower extremity fractures accounted for 97% of the injuries and a fractured 7th rib accounted for one injury (3%). Tibial fractures occurred more frequently than femoral or ankle fractures. Four (15%) fall episodes resulted in bilateral fractures. The treatment of choice was to immobilize the fractured extremity with a soft, well-padded splint. Surgical fixation was performed in only two cases. Over 80% of the patients with fall-related fractures were admitted for inpatient stays with a mean of 66 inpatient days per patient. Hospital days were most often the result of home inaccessibility, inadequate support at home, or surgical intervention.