Transcription by RNA polymerase II and the CTD-chromatin crosstalk.

The epigenetic phenomenon is known to derive the phenotypic variation of an organism through an interconnected cellular network of histone modifications, DNA methylation and RNA regulatory network. Transcription for protein coding genes is a highly regulated process and carried out by a large multi-complex RNA Polymerase II. The carboxy terminal domain (CTD) of the largest subunit of RNA Polymerase II consists of a conserved and highly repetitive heptad sequence Tyr1-Ser2-Pro3-Thr4-Ser5-Pro6-Ser7. The epigenetically modified CTD is thought to selectively bind different protein complexes that participate in mRNA biogenesis and export. The CTD and chromatin appears to have a spatial relationship during the transcription cycle, where the epigenetic modifications of CTD not only influence the state of histone modification but also mediates CTD-chromatin crosstalk. In this mini review, we have surveyed and discussed current developments of RNA Polymerase II CTD and its new emerging crosstalk with chromatin, during the stage specific progression of RNA Polymerase II in transcription cycle. This review is mainly focussed on the insights in budding yeast.

Evaluation of cytotoxic activity and genotoxicity of structurally well characterized potent cobalt(II) phen-based antitumor drug entities: An in vitro and in vivo approach.

Cobalt (II) phen-based drug candidates of the formulation Co(phen)2Cl2,1, Co(phen)2L, 2 where L = 1H-pyrazole-3,5-dicarboxylic acid were synthesized and thoroughly characterized by spectroscopic methods and single X-ray crystallography. DNA binding interaction of 1 and 2 was carried out employing biophysical techniques {UV-visible, fluorescence, thermal denaturation and cyclic voltammetry} to validate their potential to act as antitumor agents. The interpretations of these biophysical studies of 1 and 2 supported the non-covalent intercalative binding mode; furthermore, a higher binding trend of 2 was observed as compared to 1, phen and 1H-pyrazole-3,5-dicarboxylic acid alone. Cleavage studies of 1 and 2 with pBR322 were assessed by gel electrophoresis and it was observed that both drug candidates cleave DNA by hydrolytic pathway involving hydroxyl radical (OH). Cytotoxic activity of 1 and 2 against human cancer cell lines [MCF-7 (breast), HeLa (cervical), MIA-PA-CA 2 (pancreatic), A-498 (kidney), Hep-G2 (hepatoma)] was evaluated by SRB assay. The obtained results showed that drug candidate 1 showed significantly low GI50 value (<10 µg/ml) against MCF-7 and HeLa cell lines. However, candidate 2 revealed excellent cytotoxicity (<10 µg/ml) against all the tested cancer cell lines. The in vivo genotoxicity of 2 was evaluated by micronucleus (MN) test and chromosomal aberration (CA) in bone marrow cells of the Wistar rats to check cobalt(II)-induced systemic toxicity. The results showed that no significant chromosomal aberrations and micronucleus formation was observed at 5 mg/kg and 10 mg/kg in presence of drug candidate 2 implicating that it could be administered safely at a low dosage. However, an elevated percentage of chromosomal aberration and micronucleated polychromatic erythrocytes (MNPCE) was observed only at higher doses (20 mg/kg and 40 mg/kg) of drug candidate 2.

Embracing the Timeless Value of Vintage Instrumentation in Orthopedics for Broken Femoral Nail Extraction with Novel Technique.

Introduction: Intramedullary nail placement is a preferred method for femoral shaft fractures due to biomechanical advantages. Despite its simplicity and efficacy, complications, such as pseudoarthrosis and nail fracture, pose challenges. Removing the distal fragment presents a vexing issue, often requiring specialized instruments. Current retrograde techniques may not be universally accessible, potentially leading to non-union site opening and tissue damage. While intramedullary nailing benefits fracture treatment, addressing its removal complications is crucial for further advancements. In this case, universal zig and K-nail reamers are used to extract the broken nail. Case Report: A 42-year-old male suffered an injury to his left thigh in a bike collision. Radiological examination revealed an oblique fracture in the proximal 1/3rd and distal 2/3rd junction of the left femur shaft. Closed reduction and internal fixation were initially performed, but delayed union occurred. Nine months post-surgery, a broken femoral nail and non-united fracture were observed. The patient underwent successful removal and replacement of the broken nail with a wider intramedullary nail. The procedure involved using a universal zig for removal of the proximal fragment and K-nail reamers to remove the distal fragment of the broken nail, and inserting a wider proximal femoral nail. The patient was discharged in satisfactory condition with tolerable weight-bearing. Conclusion: Broken femoral nail presents a unique challenge to the operating surgeon. It requires diligent planning with appropriate instruments and techniques. There have been many techniques devised to tackle this problem and the use of K-nail reamers as described in this case is another innovative method in this regard.

Management of post-traumatic ankle deformities in children.

The objective of this clinical appraisal was to assess the clinical-radiological results of ankle deformity correction secondary to physeal injury, utilizing the methods based on the age of the child, site & severity of the deformity, remaining growth potential, condition of the soft tissue envelop and integrity of neurovascular status. Fifteen subjects ≤ 16 years of age, with angular deformities of the ankle secondary to physeal injury, were included. Deformities secondary to infection and pathological fractures were excluded. Demographic data, type of injury, treatment method, and follow-up were recorded from the case files. Treatment categories included osteotomies for acute correction (> 10 years) and growth modulation (≤ 10 years). Male to female ratio was 7:8, with an average age of 11.8 ± 2.31 years (range 9-16 years). The right and left ratio was 7:8. Mean duration of follow-up was 1 year and 4 months. Gradual deformity correction was done in 2 cases utilizing the principle of growth modulation, while acute correction by osteotomy was done in 13 cases. The average pre-operative ankle deformity was 20.8 ± 3.11 degrees (Range -25 to 24 degrees). Radiological union was attained at a mean of 11 weeks (8-24). Nine patients achieved neutral ankle alignment. The mean residual varus was 2.3°, and the valgus was 4°. There was a statistically significant improvement of the AOFAS score by 17 points from a mean pre-operative score of 57 (44-84) to 74 (56-100) points at the final follow-up (p-value < 0.001). The average pre-operative shortening was 2.36 ± 0.21 cm, which was completely corrected in 9 individuals. Management of angular deformities around the ankle calls attention to correcting the resultant angular deformity and/or limb length disparity, utilizing acute or gradual correction. A successful outcome depends on early recognition and patient-specific treatment of paediatric ankle fracture patterns. Correlating the results of our study with the available literature, we feel that both acute or gradual correction for angular deformities around the ankle is a feasible solution as long as principles of deformity correction are adhered to. Techniques for salvaging and restoring the viability of injured physeal plate warrant additional research.

Protective effect of quercetin and thymoquinone against genotoxicity and oxidative stress induced by ZnO nanoparticles in the Wistar rat model.

Zinc oxide nanoparticles (ZnO-NPs) are increasingly used in a variety of consumer and other commercial products. Hence, man faces the risk of exposure to ZnO-NPs and the consequent adverse health effects. Mitigation/prevention of such effects using natural products has drawn the attention of scientists. Therefore, the aim of the present study has been to find the toxic effects associated with exposure to ZnO-NPs, and the protective role of the phytochemicals thymoquinone (TQ) and quercetin (QCT) in the rat model. ZnO-NPs were administered to male Wistar rats through oral route; TQ / QCT was concurrently administered through intra-peritoneal route. The response in the animal was analyzed adopting chromosomal aberration test, micronucleus test, and comet assay of bone marrow cells to assess the genotoxicity, and biochemical assays of superoxide dismutase (SOD), catalase (CAT), lipid peroxidation (LPO), total extractable protein of liver, and reduced glutathione (GSH) of liver homogenate to monitor the changes in the antioxidant defense mechanism in response to the oxidative stress. Treatment of 300 mg/kg body weight (bw) of ZnO-NPs produced adverse effects on all aspects analyzed viz., structural chromosomal aberrations, micronuclei formation, DNA damage, SOD, catalase, lipid peroxidation, GSH, and extractable total protein of liver. Co-treatment of TQ / QCT offered protection against the toxicity induced by ZnO-NPs. The most optimum doses of TQ and QCT that offered the best protection were 18 mg/kg bw and 500 mg/kg bw, respectively. The study reveals that TQ / QCT supplementation is beneficial in the context of toxic effects of ZnO-NPs.

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.

Influence of zinc levels on the toxic manifestations of lead exposure among the occupationally exposed workers.

Lead and zinc are usually found at the same occupational places and occur as co-contaminants. Effects of lead toxicity are detrimental on human health as it is probable carcinogen and impairs normal growth and development. On the other hand, zinc is an important nutritional element, the deficiency of which causes debilitating effects on growth and development. The purpose of this study was to examine the possible association of blood lead and zinc levels and any influence of zinc over DNA damage, blood cell membrane aberration and oxidative stress among lead and zinc co-exposed workers. Atomic absorption spectroscopy was used for lead and zinc measurement and comet assay for DNA damage assessment. Haematological aberrations were studied using light and electron microscopy (LM and EM) followed by electron density X-ray spectroscopy (EDS) and elemental mapping. Occupational exposure was observed to cause significant elevation in blood lead levels among workers. This elevation in lead levels and associated DNA damage among workers was significantly high in comparison to controls. Further light and electron micrographs of red blood cells revealed significant morphological alterations associated with increased lead ions in workers. It was clear from SEM-based elemental maps and EDS graphs that elevated lead levels were associated with low levels of zinc. The results suggest that lead absorption is highly influenced due to zinc levels in body which has an impact over DNA damage, blood cell aberration and oxidative stress caused by lead exposure. Efforts are going on to understand the role of other trace metals on lead toxicity in order to develop a sustainably nutrition-based therapeutic intervention. Graphical abstract.

Evaluation of DNA interaction, genotoxicity and oxidative stress induced by iron oxide nanoparticles both in vitro and in vivo: attenuation by thymoquinone.

Iron oxide nanoparticles (IONPs) are known to induce cytotoxicity in various cancer cell lines through the generation of reactive oxygen species (ROS). However, the studies on its potential to induce toxicity in normal cell lines and in vivo system are limited and ambiguity still exists. Additionally, small molecules are known to interact with the DNA and cause damage to the DNA. The present study is designed to evaluate the potential interaction of IONPs with DNA along with their other toxicological effects and subsequent attenuation by thymoquinone both in vitro (primary lymphocytes) and in vivo (Wistar rats). IONPs were characterized by TEM, SEM-EDS, and XRD. The results from DNA interaction studies showed that IONPs formed a complex with DNA and also got intercalated between the base pairs of the DNA. The decrease in percent cell viability of rat's lymphocytes was observed along with an increase in ROS generation in a dose-dependent manner (50, 100, 200, 400 and 800 µg/ml of IONPs). The genetic damage in in vivo might be due to the generation of ROS as depletion in anti-enzymatic activity was observed along with an increase in lipid peroxidation in a dose-dependent manner (25, 50, 100 mg/kg of IONPs). Interestingly, supplementation of thymoquinone in combination with IONPs has significantly (P < 0.05) attenuated the genetic and oxidative damage in a dose-dependent manner both in vitro and in vivo. It can be concluded that thymoquinone has the potential to attenuate the oxidative stress and genetic toxicity in vitro and in vivo.

Protective role of nimbolide against chemotherapeutic drug hydroxyurea induced genetic and oxidative damage in an animal model.

Nimbolide is known to be an antioxidant found in neem plant. Hydroxyurea is a medication frequently used in sickle-cell disease, different cancers and HIV infection. The present study aimed to evaluate the adverse effect of HU and possible amelioration by nimbolide in Wistar rats. To test our hypothesis, we performed genotoxicity tests, biochemical assays, and histopathological studies. We observed that HU caused higher levels of genotoxicity in the treated animals. The observed genetic and oxidative damage might be due to the presence of reactive species as HU increased the level of the malondialdehyde-a biomarker of oxidative damage. Interestingly, co-treatment of animals with HU and nimbolide showed a lower level of damage. We conclude that nimbolide significantly protects the cells from the adverse effect of HU and could be considered as a potential adjuvant for the patients under HU therapy.