Divalent siRNAs are bioavailable in the lung and efficiently block SARS-CoV-2 infection.

The continuous evolution of SARS-CoV-2 variants complicates efforts to combat the ongoing pandemic, underscoring the need for a dynamic platform for the rapid development of pan-viral variant therapeutics. Oligonucleotide therapeutics are enhancing the treatment of numerous diseases with unprecedented potency, duration of effect, and safety. Through the systematic screening of hundreds of oligonucleotide sequences, we identified fully chemically stabilized siRNAs and ASOs that target regions of the SARS-CoV-2 genome conserved in all variants of concern, including delta and omicron. We successively evaluated candidates in cellular reporter assays, followed by viral inhibition in cell culture, with eventual testing of leads for in vivo antiviral activity in the lung. Previous attempts to deliver therapeutic oligonucleotides to the lung have met with only modest success. Here, we report the development of a platform for identifying and generating potent, chemically modified multimeric siRNAs bioavailable in the lung after local intranasal and intratracheal delivery. The optimized divalent siRNAs showed robust antiviral activity in human cells and mouse models of SARS-CoV-2 infection and represent a new paradigm for antiviral therapeutic development for current and future pandemics.

Bayesian optimal stepped wedge design.

Recently, there has been a growing interest in designing cluster trials using stepped wedge design (SWD). An SWD is a type of cluster-crossover design in which clusters of individuals are randomized unidirectional from a control to an intervention at certain time points. The intraclass correlation coefficient (ICC) that measures the dependency of subject within a cluster plays an important role in design and analysis of stepped wedge trials. In this paper, we discuss a Bayesian approach to address the dependency of SWD on the ICC and robust Bayesian SWDs are proposed. Bayesian design is shown to be more robust against the misspecification of the parameter values compared to the locally optimal design. Designs are obtained for the various choices of priors assigned to the ICC. A detailed sensitivity analysis is performed to assess the robustness of proposed optimal designs. The power superiority of Bayesian design against the commonly used balanced design is demonstrated numerically using hypothetical as well as real scenarios.

Cytosine methylation by DNMT2 facilitates stability and survival of HIV-1 RNA in the host cell during infection.

The enigmatic methyltransferase, DNMT2 (DNA methyltransferase 2), structurally resembles a DNA methyltransferase, but has been shown to be a tRNA methyltransferase targeting cytosine within a specific CpG in different tRNA molecules. We had previously shown that, during environmental stress conditions, DNMT2 is re-localized from the nucleus to the cytoplasmic stress granules (SGs) and is associated with RNA-processing proteins. In the present study, we show that DNMT2 binds and methylates various mRNA species in a sequence-independent manner and gets re-localized to SGs in a phosphorylation-dependent manner. Importantly, our results indicate that HIV-1 enhances its survivability in the host cell by utilizing this RNA methylation capability of DNMT2 to increase the stability of its own genome. Upon infection, DNMT2 re-localizes from the nucleus to the SGs and methylates HIV-1 RNA. This DNMT2-dependent methylation provided post-transcriptional stability to the HIV-1 RNA. Furthermore, DNMT2 overexpression increased the HIV-1 viral titre. This would suggest that HIV hijacks the RNA-processing machinery within the SGs to ensure its own survival in the host cell. Thus, our findings provide for a novel mechanism by which virus tries to modulate the host cell machinery to its own advantage.

Nanosuspension-Based Aloe vera Gel of Silver Sulfadiazine with Improved Wound Healing Activity.

The present study focuses on the development and characterization of nanosuspension of a poorly soluble drug, silver sulfadiazine (SSD) incorporated in Aloe vera gel (AV-gel) for improving its therapeutic efficacy. The SSD solution in ammonia was subjected to nanoprecipitation in surfactant solution and particle size was optimized by varying concentration of surfactant. Optimized formulation constituted of 5.5% (w/v) Span 20 and 5.5% (w/v) Tween 80 as a dispersing agent and 0.5% (w/v) Poloxamer 188 as a co-surfactant. The prepared nanosuspension was evaluated for particle size, polydispersity index, surface morphology, and x-ray diffraction study. The optimized nanosuspension was incorporated into nanogel formulation with the addition of 1% AV-gel and 0.5% Carbopol 940 for topical delivery of nanosized SSD. Evaluation of in vitro drug release exhibited a significant enhancement in release rate of the drug from developed nanogel formulation (77.16 ± 3.241%) in comparison to marketed formulation (42.81 ± 1.452%) after 48 h. In vivo histopathological studies in rats for 14 days of application of prepared nanogel showed improvement in the wound healing potential as compared to marketed formulation.

The quorum sensing molecule N-acyl homoserine lactone produced by Acinetobacter baumannii displays antibacterial and anticancer properties.

Secretory N-acyl homoserine lactones (AHLs) mediate quorum sensing (QS) in bacteria. AHLs are shown to be inhibitory for an unrelated group of bacteria and might mimic host signalling elements, thereby subverting the regulatory events in host cells. This study investigated the AHL produced by Acinetobacter baumannii and analysed its effect on other bacterial species and mammalian cells. Chemically characterized AHL had an m/z value of 325 with a molecular formula C18H31NO4 and showed its inhibitory potential against Staphylococcus aureus. Molecular docking studies identified D-alanine-D-alanine synthetase A, a cell wall synthesizing enzyme of S. aureus having a strong binding affinity towards AHL. Electron microscopy showed the disruption and sloughing off of the S. aureus cell wall when treated with AHL. In vitro experiments revealed that this bacteriostatic AHL showed time-dependent activity and induced apoptosis in cancer cell lines. This compound could be a potential structural backbone for constructing new AHL analogues against S. aureus. The findings emphasize the need to re-evaluate all previously characterized AHLs for any additional new biological functions other than QS.

Scapulothoracic and scapholunate dissociation in the ipsilateral upper limb of a trauma victim.

Scapulothoracic dissociation is a rare and complex injury pattern with varied presentation. Here we describe a case of a 32-year-old male who presented with scapulothoracic dissociation associated with brachial plexus injury, along with scapholunate dissociation. We also propose an injury mechanism that might link the two injury patterns, suggesting that the association might be more than by chance. The patient was managed according to established trauma care and resuscitation protocols followed by open reduction and internal fixation of the clavicle fracture, and fixation of scapholunate dissociation and had a successful outcome at follow-up.

OOA-modified Bi-LSTM network: An effective intrusion detection framework for IoT systems.

Currently, the Internet of Things (IoT) generates a huge amount of traffic data in communication and information technology. The diversification and integration of IoT applications and terminals make IoT vulnerable to intrusion attacks. Therefore, it is necessary to develop an efficient Intrusion Detection System (IDS) that guarantees the reliability, integrity, and security of IoT systems. The detection of intrusion is considered a challenging task because of inappropriate features existing in the input data and the slow training process. In order to address these issues, an effective meta heuristic based feature selection and deep learning techniques are developed for enhancing the IDS. The Osprey Optimization Algorithm (OOA) based feature selection is proposed for selecting the highly informative features from the input which leads to an effective differentiation among the normal and attack traffic of network. Moreover, the traditional sigmoid and tangent activation functions are replaced with the Exponential Linear Unit (ELU) activation function to propose the modified Bi-directional Long Short Term Memory (Bi-LSTM). The modified Bi-LSTM is used for classifying the types of intrusion attacks. The ELU activation function makes gradients extremely large during back-propagation and leads to faster learning. This research is analysed in three different datasets such as N-BaIoT, Canadian Institute for Cybersecurity Intrusion Detection Dataset 2017 (CICIDS-2017), and ToN-IoT datasets. The empirical investigation states that the proposed framework obtains impressive detection accuracy of 99.98 %, 99.97 % and 99.88 % on the N-BaIoT, CICIDS-2017, and ToN-IoT datasets, respectively. Compared to peer frameworks, this framework obtains high detection accuracy with better interpretability and reduced processing time.

Bilateral traumatic proximal humerus fractures managed by open reduction and internal fixation with locked plates.

Fractures of the proximal humerus are uncommon in young patients. Although bilateral fracture of proximal humerus itself is rare, association with epilepsy and electrocution is frequent. Only one case of traumatic bilateral proximal humerus fracture has been reported in the literature. We report a rare case of bilateral traumatic displaced proximal humerus fractures in a 40 years old male patient, which was treated by means of open reduction and internal fixation with proximal humerus locked pates on both sides and obtained a good functional outcome.

Prognostic value of Ocular Trauma Score and pediatric Penetrating Ocular Trauma Score in predicting the visual prognosis following ocular injury.

Aim: To assess the role of Ocular Trauma Score (OTS) and Pediatric Penetrating Ocular Trauma Score (POTS) in predicting visual prognosis following ocular injury. Methods: 120 patients, aged 3 to 70 years, with ocular injury, presenting between August 2020 and 2021, who fulfilled the inclusion criteria, were classified using the Birmingham Eye Trauma Terminology System (BETTS). Data regarding age, sex, injury type, location, initial visual acuity, and treatment were recorded. Patients were evaluated using OTS and POTS, wherever applicable, to predict visual prognosis. Follow-up was done for 6 months post-treatment to compare the predicted and actual visual outcome. Results: 120 patients (96 males and 24 females) were included in the study. The mean age was 17.2 ± 14.7 years (range 3 to 70 years). 91.6% patients included were under the age of 40 years. Blunt trauma (n=62) was slightly more common. 80 patients (66.67%) reported to the hospital after 48 hours of injury. 34 patients (28.33%) had traumatic cataract alone and IOL implantation was done as a primary procedure. In 20 patients (16.67%), lens aspiration was done primarily, with secondary IOL implantation. Six months after the treatment, the BCVA in the open globe injury patients was 20/ 200 or better in 36 patients (75%) and 20/ 40 or better in 18 patients (37.5%). Amongst the closed globe injuries, 48 patients (66.67%), had a BCVA 20/ 200 or better, while 32 (44.44%) had a BCVA of 20/ 40 or better. Conclusion: OTS is a reliable predictor of final visual outcome, both in blunt and penetrating injuries. The POTS shows gross underestimation of final visual outcome. Abbreviations: BETTS = Birmingham Eye Trauma Terminology System, POTS = Pediatric Penetrating Ocular Trauma Score, OTS = Ocular Trauma Score, OGI = Open Globe Injury, HM = Hand Movement, NLP = No Light Perception, LP = Light Perception, CGI = Closed Globe Injury, VA = Visual Acuity, BCVA = Best Corrected Visual Acuity, IAP = Indian Academy of Pediatrics, IOL = Intraocular Lens, IGATES = International Globe and Adnexal Trauma Epidemiology Study, IOP = Intraocular Pressure, CART = Classification and Regression Tree, USA = United States of America.

Optimal allocation of subjects in a matched pair cluster-randomized trial with fixed number of heterogeneous clusters.

In cluster-randomized trials, investigators randomize clusters of individuals such as households, medical practices, schools or classrooms despite the unit of interest are the individuals. It results in the loss of efficiency in terms of the estimation of the unknown parameters as well as the power of the test for testing the treatment effects. To recoup this efficiency loss, some studies pair similar clusters and randomize treatment within pairs. However, the clusters within a treatment arm might be heterogeneous in nature. In this article, we propose a locally optimal design that accounts the clusters heterogeneity and optimally allocates the subjects within each cluster. To address the dependency of design on the unknown parameters, we also discuss Bayesian optimal designs. Performances of proposed designs are investigated numerically through some data examples.

Rhino-orbital mucormycosis: Our experiences with clinical features and management in a tertiary care center.

Objective: To determine the prevalence, risk factors, and elaborate our experiences with diagnosis and treatment of patients with mucormycosis, enabling a better understanding of the disease and its management. Methods: This is a case series of patients with Covid-19 associated with Rhino-orbital-cerebral mucormycosis, managed in our tertiary care center from April 2021 to June 2021. Results: Six cases of Covid-19 associated with Rhino-orbital-cerebral mucormycosis have been analyzed in the study. The mean age of patients was 40.67 years with a male preponderance (83.3%). The most common complaint was headache (100%), while a minority (33%) came with ocular complaints. All the patients either had a previous history of diabetes mellitus or developed increased blood sugar levels following Covid infection, and were kept on insulin to control their blood sugar levels. 4 patients (66.67%) had a history of corticosteroid use during Covid-19 hospitalization. Treatment included intravenous liposomal Amphotericin B (100%), functional endoscopic sinus surgery (66.67%), maxillectomy (33.33%) and transcutaneous retrobulbar liposomal Amphotericin B (33.33%). Amphotericin B induced nephrotoxicity, which was seen in 1 patient (16.67%). Mortality occurred in only one patient (16.67%), 25 days following successful surgery. Conclusion: Diabetes Mellitus is the most important predisposing factor for the development of Covid-19 associated Rhino-orbital-cerebral mucormycosis. Early presentation, prompt diagnosis and timely initiation of treatment with liposomal Amphotericin B and surgical debridement along with strict blood sugar control can lead to a favorable outcome. However, regular follow-up and monitoring of serum electrolytes and kidney profile must be ensured for such patients.

Viral protein X unlocks the nuclear pore complex through a human Nup153-dependent pathway to promote nuclear translocation of the lentiviral genome.

Simian immunodeficiency virus (SIV) and human immunodeficiency virus 2 (HIV-2) display unique ability to infect nondividing target cells. Viral protein X (Vpx) of HIV-2/SIV is known to be involved in the nuclear import of viral genome in nondividing cells, but the mechanism remains poorly understood. In the present investigation for the first time we provide evidence that Vpx of SIVsmPBj1.9 physically interacts with human nucleoporin 153 (Nup153), which is known to provide a docking site for protein-cargo complexes at the nuclear pore complex (NPC). Results from superresolution-structured illumination microscopy studies reveal that Vpx interaction with NPC-associated Nup153 is critical for its efficient nuclear translocation. Virion-associated MAPK/ERK-2-mediated phosphorylation of Vpx plays a critical role in its interaction with human Nup153 and this interaction was found to be evolutionarily conserved in various SIV isolates and HIV-2. Interestingly, MAPK/ERK-2 packaging defective SIV failed to promote the efficient nuclear import of viral genome and suggests that MAPK/ERK-2-mediated Vpx phosphorylation is important for its interaction with Nup153, which is critical for lentiviruses to establish infection in nondividing target cells. Together, our data elucidate the mechanism by which Vpx orchestrates the challenging task of nuclear translocation of HIV-2/SIV genome in nondividing target cells.

Bioinspired Smart Nanosystems in Advanced Therapeutic Applications.

BACKGROUND: Nanoparticle technologies used for human administration must be designed to interact with a living host environment. The idea about bioinspired smart drug delivery carriers includes the development of biocompatible nanomaterials which can be further loaded with the drug for specific targeted drug delivery applications. OBJECTIVE: Biosmart nanosystems are used for several applications in the delivery of drugs and pharmaceuticals for their therapeutic applications like biological markers, diagnostic purposes such as imaging applications and also for gene therapy. Thus, the bioinspired nanocarriers are capable of carrying biologically active molecules to the target sites. This bioinspired nanosystem constitutes of lipids, polymers and biomaterials which utilizes various responsive sensors for targeted drug delivery systems. However, external conditions such as heat, light, magnetic or electric field and ultrasounds, along with temperature, altered pH and ionic strength can affect the bioinspired smart nanosystem for drug delivery. CONCLUSION: The present review focuses on challenges for the development of bioinspired smart nanocarriers for the management of various disorders.

An Overview on Genistein and its Various Formulations.

Genistein is the natural isoflavone and a phytoestrogen with a broad range of pharmacological properties, such as tyrosine and topoisomerase inhibition. It also induces apoptosis and cell proliferation inhibition, differentiates cancer cells. Added health benefits include the reduction of osteoporosis by suppressing osteoclasts and lymphocyte functions, decreased the risk of cardiovascular attacks and relieved postmenopausal problems. Genistein traditionally used in Chinese and Ayurvedic medicine and are found to be associated with lower risk of breast, prostate and lung cancer. Numerous factors comprising genetic, epigenetic and transcriptomic alterations are evidenced to be responsible for breast, prostate and lung cancer. In present review, an overview on genistein, the various analytical methods and drug delivery approaches to determine genistein in the formulations are discussed. It may help to develop novel formulations with better solubility and bioavailability of genistein. The tumor cell scan may be targeted to form a stable genistein formulation.

Therapeutic Intervention of Aloe Gel Containing Nano-Sized and Micron-Sized Silver Sulfadiazine Gel on Second-Degree Burn: A Comparative Study.

The current work focuses on the formulation development, optimization, and in vivo assessment of nano-sized silver sulfadiazine ( nSSD) and micron-sized silver sulfadiazine ( mSSD) topical gel composed of Aloe vera gel ( Aloe gel) and Carbopol 940 for the management of second-degree burn wound. The optimized concentration of gel-forming agent (Carbopol 940) was chosen based on best possible consistency and spreadability of the gel. The second-degree burn infliction was developed in the posterior region of rats followed by anesthesia. Afterward, the created wounds were further treated individually by both the gel formulation (1 application daily) for 14 days and observations were recorded. The nSSD gel showed better wound healing and a higher degree of tissue hyperplasia as compared with mSSD gel in rats. In vitro drug release study showed better drug release from nSSD gel (74.25 ± 3.331%) as compared with mSSD gel formulation (61.32 ± 2.112%) after 24 hours. The nSSD and mSSD topical gel-treated rats showed 95.63% and 78.75% wound healing after 14 days, while in the case of control group rats, 48.65% wound contraction was seen after 14 days. Furthermore, the histopathological study revealed that the nSSD gel was more efficient in controlling the wound infection and showed better wound healing as compared with mSSD gel formulation.

Silymarin nanoemulsion against human hepatocellular carcinoma: development and optimization.

OBJECTIVE: Nanoemulsion of silymarin was developed and optimized. MATERIALS AND METHODS: Nanoemulsion was made by aqueous titration method. Sefsol 218 (5.8% v/v), Kolliphor RH40 and polyethylene glycol 400 (Smix; 2:1; 28.99% v/v) were used as oil phase, surfactant and co-surfactant while distilled water (65.22% v/v) acted as an aqueous phase. Nanoemulsion was characterized on the basis of particle size, viscosity, electrical conductivity and refractive index. Further, in vitro release, in vivo pharmacokinetic study, stability study and cancer cell line studies were also performed. RESULTS AND DISCUSSION: The optimized formulation (NE9) with mean particle size of 21.24 nm showed a minimum viscosity of 9.59 cps, maximum drug release (97.75%) in 24 h. The NE9 formulation also showed higher AUC (p < .01) and Cmax (p < .01) and shorter Tmax (p < .05) compared with conventional and standard suspensions of silymarin. The stability study also showed considerably stable formulations at refrigerator temperature as compared with room temperature (p > .05). The cancer cell line studies also confirmed that silymarin nanoemulsion reduced the cell viability and increased ROS intensity and chromatin condensation (p < .05). CONCLUSION: Our results concluded that nanoemulsion may be an efficient carrier for oral delivery of silymarin against human hepatocellular carcinoma without damaging normal cells.

Solvent Extraction and GC-MS Analysis of Sesame Seeds for Determination of Bioactive Antioxidant Fatty Acid/Fatty Oil Components.

The seed kernels of Sesamum indicum L. (family: Pedaliaceae) were extracted with ethanol and yield of components determined by Gas Chromatography/Mass Spectrometry (GC/MS). The free radical scavenging activities of ethanolic extract against1, 1-Diphenyl-2-picrylhydrazyl (DPPH) were determined by UV spectrophotometer at 517 nm. Phytochemical screening revealed the presence of numerous bioactive compounds including steroids, phenolic, terpenoids, fatty acids and different types of ester compounds. The ethanolic extract was purified and analyzed by GC MS.The prevailing compounds found in ethanolic extract were Carvacrol (0.04%),Sesamol (0.11%), 4-Allyl-2-methoxy-phenol(0.04%),Palmitic acid (1.08%), cis-9-Hexadecenal (85.40%), Lineoleoyl chloride (0.52%), Palmitic acid ß-monoglyceride (0.40%), Dihydro-aplotaxene (0.61%), Oleoyl chloride (1.11%), (+)-Sesamin (4.73%), 1,3-Benzodioxole, 5-[4-(1,3-benzodioxol-5-yloxy)tetrahydro-1 H,3 H-furo [3,4-c]furan-1-yl], [1 S-(1,3,4,6α.), (2.01%)], 6-Nitrocholest-5-en-3-yl acetate (0.22%), Ergost-5-en-3ß-ol (2.35%) and 24-Propylidenecholesterol (0.16%). The presence ofsaturated and unsaturated fatty acids in ethanolicextract justifies the use of this plant to treat many ailments in folk and traditional medicine. Ethanolic extract have shown significant antioxidant activity(IC50120.38±2.8 µg/ml). The presence of phenolic (Sesamol), lignin (Sesamin) compounds and unsaturated fatty acids are reported as possible contributor for antioxidantactivity of seed extract.

Strategies in Development and Delivery of Nanotechnology Based Cosmetic Products.

The science of formulation involving cosmetic ingredients has always been a challenge since the release of active components greatly depends upon the carrier system involved and the selectivity of skin barrier. The principle obstacle of the skin resides in the epidermis and it's hard for many active components to cross it. The formulation related factors like size of particles, viscosity and lipophilicity of the components also play an important role in permeation of the dermal composition. Though widely used; conventional creams and gels still struggle in terms of success. This work focuses on nano based formulation strategies for successful delivery of cosmetic agents. Novel strategies like nanoemulsion, nanogels, liposomes, aquasomes, niosomes, dendrimers and fullerenes have paved way for successful delivery of dermal formulations to desire depths in the skin.

Design and evaluation of sustained release bilayer tablets of propranolol hydrochloride.

The objective of the present research was to develop a bilayer tablet of propranolol hydrochloride using superdisintegrant sodium starch glycolate for the fast release layer and water immiscible polymers such as ethylcellulose, Eudragit RLPO and Eudragit RSPO for the sustaining layer. In vitro dissolution studies were carried out in a USP 24 apparatus I. The formulations gave an initial burst effect to provide the loading dose of the drug followed by sustained release for 12 h from the sustaining layer of matrix embedded tablets. In vitro dissolution kinetics followed the Higuchi model via a non-Fickian diffusion controlled release mechanism after the initial burst release. FT-IR studies revealed that there was no interaction between the drug and polymers used in the study. Statistical analysis (ANOVA) showed no significant difference in the cumulative amount of drug release after 15 min, but significant difference (p < 0.05) in the amount of drug released after 12 h from optimized formulations was observed.