Transcriptional activation of the Mycobacterium tuberculosis virulence-associated small RNA MTS1338 by the response regulators DosR and PhoP.

MTS1338, a distinctive small RNA in pathogenic mycobacteria, plays a crucial role in host-pathogen interactions during infection. Mycobacterial cells encounter heterogeneous stresses in macrophages, which highly upregulate MTS1338. A dormancy regulatory factor DosR regulates the intracellular abundance of MTS1338. Herein, we investigated the interplay of DosR and a low pH-inducible gene regulator PhoP binding to the MTS1338 promoter. We identified that DosR strongly binds to two regions upstream of the MTS1338 gene. The proximal region possesses a threefold higher affinity than the distal site, but the presence of both regions increased the affinity for DosR by > 10-fold. PhoP did not bind to the MTS1338 gene but binds to the DosR-bound MTS1338 gene, suggesting a concerted mechanism for MTS1338 expression.

A novel ionizing radiation-induced small RNA, DrsS, promotes the detoxification of reactive oxygen species in Deinococcus radiodurans.

A plethora of gene regulatory mechanisms with eccentric attributes in Deinoccocus radiodurans confer it to possess a distinctive ability to survive under ionizing radiation. Among the many regulatory processes, small RNA (sRNA)-mediated regulation of gene expression is prevalent in bacteria but barely investigated in D. radiodurans. In the current study, we identified a novel sRNA, DrsS, through RNA-seq analysis in D. radiodurans cells while exposed to ionizing radiation. Initial sequence analysis for promoter identification revealed that drsS is potentially co-transcribed with sodA and dr_1280 from a single operon. Elimination of the drsS allele in D. radiodurans chromosome resulted in an impaired growth phenotype under γ-radiation. DrsS has also been found to be upregulated under oxidative and genotoxic stresses. Deletion of the drsS gene resulted in the depletion of intracellular concentration of both Mn2+ and Fe2+ by ~70% and 40%, respectively, with a concomitant increase in carbonylation of intracellular protein. Complementation of drsS gene in ΔdrsS cells helped revert its intracellular Mn2+ and Fe2+ concentration and alleviated carbonylation of intracellular proteins. Cells with deleted drsS gene exhibited higher sensitivity to oxidative stress than wild-type cells. Extrachromosomally expressed drsS in ΔdrsS cells retrieved its oxidative stress resistance properties by catalase-mediated detoxification of reactive oxygen species (ROS). In vitro binding assays indicated that DsrS directly interacts with the coding region of the katA transcript, thus possibly protecting it from cellular endonucleases in vivo. This study identified a novel small RNA DrsS and investigated its function under oxidative stress in D. radiodurans. IMPORTANCE: Deinococcus radiodurans possesses an idiosyncratic quality to survive under extreme ionizing radiation and, thus, has evolved with diverse mechanisms which promote the mending of intracellular damages caused by ionizing radiation. As sRNAs play a pivotal role in modulating gene expression to adapt to altered conditions and have been delineated to participate in almost all physiological processes, understanding the regulatory mechanism of sRNAs will unearth many pathways that lead to radioresistance in D. radiodurans. In that direction, DrsS has been identified to be a γ-radiation-induced sRNA, which is also induced by oxidative and genotoxic stresses. DrsS appeared to activate catalase under oxidative stress and detoxify intracellular ROS. This sRNA has also been shown to balance intracellular Mn(II) and Fe concentrations protecting intracellular proteins from carbonylation. This novel mechanism of DrsS identified in D. radiodurans adds substantially to our knowledge of how this bacterium exploits sRNA for its survival under stresses.

Selective Bacterial Growth Inactivation by pH-Sensitive Sulfanilamide Functionalized Carbon Dots.

Carbon dots (CDs) were synthesized hydrothermally by mixing citric acid (CA) and an antifolic agent, sulfanilamide (SNM), employed for pH sensing and bacterial growth inactivation. Sulfanilamide is a prodrug; aromatic hetero cyclization of the amine moiety along with other chemical modifications produces an active pharmacological compound (chloromycetin and miconazole), mostly administered for the treatment of various microbial infections. On the other hand, the efficacy of the sulfanilamide molecule as a drug for antimicrobial activity was very low. We anticipated that the binding of the sulfanilamide molecule on the carbon dot (CD) surface may form antibacterial CDs. Citric acid was hybridized with sulfanilamide during the hydrothermal preparation of the CDs. The molecular fragments of bioactivated sulfanilamide molecule play a crucial role in bacterial growth inactivation for Gram-positive and Gram-negative bacteria. The functional groups of citric acid and sulfanilamide were conserved during the CD formation, facilitating the zwitterionic behavior of CDs associated with its photophysical activity. At low concentrations of CDs, the antibacterial activity was apparent for Gram-positive bacteria only. This Gram-positive bacteria selectivity was also rationalized by zeta potential measurement.

Is salvage Plug-Assisted Retrograde Transvenous Obliteration (PARTO) safe and effective for bleeding gastric varices ?- A preliminary single-center experience.

BACKGROUND: Vascular plug-assisted retrograde transvenous obliteration (PARTO) obliterates the gastric varices and portosystemic shunt, thus resulting in a lower rebleeding rate than endoscopic glue/sclerotherapy. AIMS: To evaluate the safety and efficacy of PARTO as salvage therapy in liver cirrhosis with gastric variceal bleed (GVB) after failed endotherapy. We assessed the clinical success rate and changes in liver function at 6- months. MATERIALS AND METHODS: Patients who underwent salvage PARTO after failed endotherapy for GVB (between December 2021 and November 2022) were searched and analyzed from the hospital database. Clinical success rate and rebleed rate were obtained at six months. Child-Pugh score (CTP) and Model for end-stage liver disease (MELD) score were calculated and compared between baseline and 6-month follow-up. RESULTS: Fourteen patients (n = 14, Child-Pugh class A/B) underwent salvage PARTO. Nine had GOV-2, and five had IGV-1 varices. The mean shunt diameter was 11.6 ± 1.6 mm. The clinical success rate of PARTO was 100% (no recurrent gastric variceal hemorrhage within six months). No significant deterioration in CTP (6.79 ± 0.98 vs. 6.21 ± 1.52; p = 0.12) and MELD scores (11.5 ± 4.05 vs. 10.21 ± 3.19; p = 0.36) was noted at 6 months. All patients were alive at 6 months. One patient (n = 1, 7.1%) bled from esophageal varices after three days of PARTO and was managed with variceal banding. 21.4% (3/14) patients had progression of esophageal varices at 6 months requiring prophylactic band ligation. Three patients (21.4%) had new onset or worsening ascites and responded to low-dose diuretics therapy. CONCLUSIONS: PARTO is a safe and effective procedure for bleeding gastric varices without any deterioration in liver function even after six months. Patient selection is critical to prevent complications. Further prospective studies with larger sample size are required to validate our findings.

Bloody Bile and Rescue Intervention-A Case Series of Post-PTBD Hemorrhagic Complications With a Review of the Literature.

Percutaneous transhepatic biliary drainage (PTBD) is a routinely performed interventional radiological procedure. A myriad of complications can occur after PTBD, the most important being hemorrhagic complications that require immediate attention. Hemorrhage following PTBD may result from arterial, portal, or hepatic venous injury. A catheter or pull-back cholangiogram often demonstrates the venous injury. A computed tomogram angiogram aids in identifying bleeding sources and procedural planning. Catheter repositioning, upsizing, or clamping often suffice for minor venous bleeding. However, major venous injury necessitates tract embolization, portal vein embolization, or stent grafting. Arterial injury may lead to significant blood loss unless treated expeditiously. Transarterial embolization is the treatment of choice in such cases. Adequate knowledge about the hemorrhagic complications of PTBD will allow an interventional radiologist to take necessary precautionary measures to reduce their incidence and take appropriate steps in their management. This article entails four different hemorrhagic complications of PTBD and their interventional management. It also discusses the various treatment options to manage different kinds of post-PTBD hemorrhagic complications.

Underwater Adhesives from Redox-Responsive Polyplexes of Thiolated Polyamide Polyelectrolytes.

We report the fabrication of optically clear underwater adhesives using polyplexes of oppositely charged partially-thiolated polyamide polyelectrolytes (TPEs). The thiol content of the constituent PEs was varied to assess its influence on the adhesive properties of the resulting glues. These catechol-free, redox-responsive TPE-adhesives were formulated in aquo and exhibited high optical transparency and strong adhesion even on submerged or moist surfaces of diverse polar substrates such as glass, aluminium, wood, and bone pieces. The adhesives could be cured under water through oxidative disulphide crosslinking of the constituent TPEs. The polyamide backbone provided multi-site H-bonding interactions with the substrates while the disulphide crosslinking provided the cohesive strength to the glue. Strong adhesion of mammalian bones (load bearing capacity upto 7 kg/cm2 ) was achieved using the adhesive containing 30 mol % thiol residues. Higher pH and use of oxidants such as povidone-iodine solution enhanced the curing rate of the adhesives, and so did the use of Tris buffer instead of Phosphate buffer. The porous architecture of the adhesive and its progressive degradation in aqueous medium over the course of three weeks bode well for diverse biomedical applications where temporary adhesion of tissues is required.

Role of Interventional Radiology (IR) in vascular emergencies among cirrhotic patients.

Gastrointestinal hemorrhage remains one of the most common causes of morbidity and mortality among patients with liver cirrhosis. Mostly, these patients bleed from the gastroesophageal varices. However, nonvariceal bleeding is also more likely to occur in these patients. Because of frequent co-existing coagulopathy, cirrhotics are more prone to bleed from a minor vascular injury while performing percutaneous interventions. Ultrasound-guided bedside vascular access is an essential procedure in liver critical care units. Transjugular portosystemic shunts (TIPS) with/without variceal embolization is a life-saving measure in patients with refractory variceal bleeding. Whenever feasible, balloon-assisted retrograde transvenous obliteration (BRTO) is an alternative to TIPS in managing gastric variceal bleeding, but without a risk of hepatic encephalopathy. In cases of failed or unfeasible endotherapy, transarterial embolization using various embolic agents remains the cornerstone therapy in patients with nonvariceal bleeding such as ruptured hepatocellular carcinoma, gastroduodenal ulcer bleeding, and procedure-related hemorrhagic complications. Among various embolic agents, N-butyl cyanoacrylate (NBCA) enables better vascular occlusion in cirrhotics, even in coagulopathy, making it a more suitable embolic agent in an expert hand. This article briefly entails the different interventional radiological procedures in vascular emergencies among patients with liver cirrhosis.

Thermostable Bacterial Laccase: Catalytic Properties and Its Application in Biotransformation of Emerging Pollutants.

Laccases have been predominantly reported in fungi, and primarily, fungal laccases are currently exploited in industrial applications. However, extremophilic bacterial laccases possess immense potential, as they can withstand extreme temperatures, pH, and salt concentrations. In addition, unlike fungal laccases, the production of bacterial laccases is cost-effective. Therefore, bacterial laccases are gaining significant attention for their large-scale applications. Previously, we reported a novel thermostable laccase (LacT) from Brevibacillus agri. Herein, we have confirmed that LacT shares a high sequence similarity with CotA laccase from Bacillus amyloliquefaciens. Peptide mass fingerprinting of LacT was conducted via matrix-assisted laser desorption ionization-time-of-flight mass spectrometry (MALDI-TOF/MS-MS). Inductively coupled plasma-optical emission spectroscopic (ICP-OES) analysis revealed the presence of ∼3.95 copper ions per protein molecule. Moreover, the secondary and tertiary structure of LacT was studied using circular dichroism (CD) and fluorescence spectroscopy. The absence of notable shifts in CD and fluorescence spectra with an increase in temperature established that LacT remains intact even at elevated temperatures. Analysis of the thermal denaturation profile of LacT by thermogravimetric analysis (TGA) also confirmed its temperature stability. Thereafter, we exploited LacT in its application for the bioremediation of phenolic endocrine disruptors, namely, triclosan, 4,4'-dihydroxybiphenyl, and dienestrol. LacT oxidizes 4,4'-dihydroxybiphenyl and triclosan but no LacT activity was detected with dienestrol. The rate of biotransformation of 4,4'-dihydroxybiphenyl and triclosan increased in the presence of CuSO4 and a redox mediator, ABTS. Transformation of dienestrol was observed only with LacT in the presence of ABTS. This study establishes the application of LacT for the bioremediation of phenolic compounds.

Combined Bile Duct Ablation and Fistulous Tract Embolization Using N-Butyl Cyanoacrylate to Manage a Biliary-Cutaneous Fistula Following Percutaneous Transhepatic Biliary Drainage (PTBD) for Hilar Cholangiocarcinoma.

Biliary fistula and bile leak are known complications following hepatobiliary surgery, trauma, and percutaneous biliary interventions. In the case of an isolated biliary system with a prolonged indwelling percutaneous transhepatic biliary drainage (PTBD) catheter, a biliary-cutaneous fistula (BCF) may develop after catheter blockage or its accidental slippage. Due to the absence of internal drainage, secreted bile flows through the matured PTBD tract to form a fistula. If left untreated, chronic BCF will result in malabsorption, infection, and delayed wound healing. Here, we report a case of left-sided BCF following prolonged PTBD for Bismuth type II cholangiocarcinoma (metastatic disease), which was initially managed by bile duct ablation using N-butyl cyanoacrylate. The patient further needed fistulous tract embolization to obliterate the BCF.

Patterning at the Resolution Limit of Commercial Electron Beam Lithography.

It has been long known that low molecular weight resists can achieve a very high resolution, theoretically close to the probe diameter of the electron beam lithography (EBL) system. Despite technological improvements in EBL systems, the advances in resists have lagged behind. Here we demonstrate that a low-molecular-mass single-source precursor resist (based on cadmium(II) ethylxanthate complexed with pyridine) is capable of a achieving resolution (4 nm) that closely matches the measured probe diameter (∼3.8 nm). Energetic electrons enable the top-down radiolysis of the resist, while they provide the energy to construct the functional material from the bottom-up─unit cell by unit cell. Since this occurs only within the volume of resist exposed to primary electrons, the minimum size of the patterned features is close to the beam diameter. We speculate that angstrom-scale patterning of functional materials is possible with single-source precursor resists using an aberration-corrected electron beam writer with a spot size of ∼1 Å.

Thermostable bacterial laccase for sustainable dyeing using plant phenols.

Laccase is regarded as an efficacious eco-friendly enzyme in various industries. Thus, various laccases have been explored to mitigate the environmental effects of conventional industrial processing; however, the prospects of laccase in hair dyeing have not been thoroughly explored to date. On account of the adverse environmental and health-related issues posed by chemical hair dyeing, laccase as a natural alternative in dyeing hair has recently gained attention. In this study, we executed hair dyeing with different colours and shades of hair dyes developed from natural plant phenols, including ferulic acid, gallic acid, catechol, and syringaldehyde, catalysed by a novel thermostable bacterial laccase (LacT) from Brevibacillus agri. The dyed hair was characterised in terms of its colourimetric parameters (L*, a*, and b*), colour strength (K/S), reflectance (R) and colour durability. L* means luminosity and is defined by L* values from 0 (black) to 100 (white). A positive value of a* means red shades and a negative value indicates green shades. A positive value of b* shows yellow shades and a negative value indicates blue shades. Optical microscopy of circular and longitudinal sections of the dyed hair revealed that the laccase-catalysed dyes did not merely stick to the surface; instead, they well-penetrated the hair. Furthermore, the dyeing process did not affect the surface morphology of the dyed hair. The dyed hair also exhibited a desirable range of colour diversity in terms of market-driven demands and showed considerable resistance to fading during shampooing and pH alterations. Post-dyeing, the texture and tensile strength of the dyed hair remained nearly unchanged. Overall, the outcomes suggest that LacT holds high potential to be exploited extensively in the hair dyeing industry as an alternative to chemical hair dyes.

Tuning the Coexistence Regime of Incomplete and Tubular Skyrmions in Ferromagnetic/Ferrimagnetic/Ferromagnetic Trilayers.

The development of skyrmionic devices requires a suitable tuning of material parameters to stabilize skyrmions and control their density. It has been demonstrated recently that different skyrmion types can be simultaneously stabilized at room temperature in heterostructures involving ferromagnets, ferrimagnets, and heavy metals, offering a new platform of coding binary information in the type of skyrmion instead of the presence/absence of skyrmions. Here, we tune the energy landscape of the two skyrmion types in such heterostructures by engineering the geometrical and material parameters of the individual layers. We find that a fine adjustment of the ferromagnetic layer thickness, and thus its magnetic anisotropy, allows the trilayer system to support either one of the skyrmion types or the coexistence of both and with varying densities.

Laccases: Thriving the domain of bio-electrocatalysis.

Laccases have been in the spotlight due to its capability to catalyse electrocatalytic reactions. The Its ability to reduce water to oxygen has made it more bio-compatible than majority of electrocatalysts. The exploitation of such biocatalysts via protein engineering and biotechnological techniques have rendered a significant breakthrough in the field of electrocatalysis. This review aims to provide a comprehensive overview of the various laccase-driven electrocatalytic reactions. Structural and functional features of laccases that contribute towards the electron transfer mechanism and reduction potential have been meticulously described. Tailoring of laccase affords an excellent prospect for its application in diverse fields like, biocathode fabrication for enzymatic biofuel cells, biosensors, and electrocatalytic reactions like water splitting. The substantial knowledge of enzyme engineering, primarily through site directed mutagenesis is propitious for the design of optimal bioelectrocatalyst. This review seeks to provide an insight for the optimisation of electrocatalytic properties of laccase thereby broadening the horizon for the development of resilient and biocompatible bioelectrocatalyst.

Synthesis and characterization of zinc derivatized 3, 5-dihydroxy 4', 7-dimethoxyflavone and its anti leishmaniasis activity against Leishmania donovani.

This study reports the synthesis and characterization of zinc derivatized 3,5-dihydroxy 4', 7- dimethoxyflavone (DHDM-Zn) compound for the development of new antileishmanial agents. The interaction studies of DHDM with zinc were carried out by UV spectra and fluorescence spectra analysis. Characterization of the complex was further accomplished by multi-spectroscopic techniques such as FTIR, Raman, HRMS, NMR, FESEM-EDX. The morphological and topographical studies of synthesized DHDM-Zn were carried out using FESEM with EDX. Further, it was demonstrated that DHDM-Zn exhibited an excellent in vitro antagonistic effect against the promastigote form of L. donovani. In addition, the possible mechanisms of promastigote L. donovani cell death, by involvement of derivatized compound in arrest of the cell cycle in the G1 phase and residual cell count reduction were investigated. Promastigote growth kinetics performed in the presence of the derivatized compound revealed a slow growth rate. The combination of growth kinetics and cell cycle analysis, made it possible to interpret and classify the cause of leishmanial cell death accurately. These results support that zinc derivatized complex (DHDM-Zn) might work as a lead compound for designing and developing a new antileishmanial drug.

Recent Advances in the Development of Oxidoreductase-Based Biosensors for Detection of Phenolic Antioxidants in Food and Beverages.

Antioxidants are known to exhibit a protective effect against reactive oxygen species (ROS)-related oxidative damage. As a result, inclusion of exogenous antioxidants in the diet has greatly increased. In this sense, detection and quantification of such antioxidants in various food and beverage items are of eminent importance. Monophenols and polyphenols are among the most prominent natural antioxidants. In this regard, biosensors have emerged as a simple, fast, and economical method for determination of such antioxidants. Owing to the fact that majority of the phenolic antioxidants are electroactive, oxidoreductase enzymes are the most extensively availed bioreceptors for their detection. Herein, the different types of oxidoreductases that have been utilized in biosensors for the biorecognition and quantification of natural phenolic compounds commonly present in foods and beverages are discussed. Apart from the most accustomed electrochemical biosensors, this review sheds light on the alternative transduction systems for the detection of phenolic antioxidants. Recent advances in the strategies involved in enzyme immobilization and surface modification of the biosensing platform are analyzed. This review aims to provide a brief overview of the latest developments in biosensor technology for phenolic antioxidant analysis in foodstuffs and future directions in this field.

PhoP induces RyjB expression under acid stress in Escherichia coli.

Bacterial small RNAs (sRNAs) play a pivotal role in post-transcriptional regulation of gene expression and participate in many physiological circuits. An ~80-nt-long RyjB was earlier identified as a novel sRNA, which appeared to be accumulated in all phases of growth in Escherichia coli. We have taken a comprehensive approach in the current study to understand the regulation of ryjB expression under normal and pH stress conditions. RpoS was not necessary for ryjB expression neither at normal condition nor under acid stress. Hfq also emerged to be unnecessary for RyjB accumulation. Interestingly, RyjB was detected as a novel acid stress induced sRNA. A DNA binding protein PhoP, a component of PhoP/Q regulon, was found to regulate ryjB expression at low pH, as the elimination of phoP allele in the chromosome exhibited a basal level of RyjB expression under acid stress. Ectopic expression of PhoP in ΔphoP cells restored the overabundance of RyjB in the cell. Overexpression of RyjB increased the abundance of sgcA transcripts, with which RyjB shares a 4-nt overlap. The current study increases our knowledge substantially regarding the regulation of ryjB expression in E. coli cell.

MTS1338 in Mycobacterium tuberculosis promotes detoxification of reactive oxygen species under oxidative stress.

Diverse mechanisms exist in Mycobacterium tuberculosis for adaptation to stresses leading to its persistence in the hostile environment of macrophages. Small RNA (sRNA)-mediated regulatory mechanisms have been scarcely explored in M. tuberculosis. MTS1338, a sRNA present only in pathogenic mycobacteria, was discovered to be highly abundant during infection and significantly contributes to host-pathogen interaction. A variety of stresses have been implicated for its accumulation. Herein, we showed that MTS1338 is an oxidative stress induced sRNA, which promotes the detoxification of reactive oxygen species (ROS) under oxidative stress. Current study identified a new role of MTS1338 in M. tuberculosis under oxidative stress.