Anticancer therapeutic potential of multimodal targeting agent- "phosphorylated galactosylated chitosan coated magnetic nanoparticles" against N-nitrosodiethylamine-induced hepatocellular carcinoma.

Superparamagnetic iron oxide nanoparticles (SPIONs) are extensively used as carriers in targeted drug delivery and has several advantages in the field of magnetic hyperthermia, chemodynamic therapy and magnet assisted radionuclide therapy. The characteristics of SPIONs can be tailored to deliver drugs into tumor via "passive targeting" and they can also be coated with tissue-specific agents to enhance tumor uptake via "active targeting". In our earlier studies, we developed HCC specific targeting agent- "phosphorylated galactosylated chitosan"(PGC) for targeting asialoglycoprotein receptors. Considering their encouraging results, in this study we developed a multifunctional targeting system- "phosphorylated galactosylated chitosan-coated magnetic nanoparticles"(PGCMNPs) for targeting HCC. PGCMNPs were synthesized by co-precipitation method and characterized by DLS, XRD, TEM, VSM, elemental analysis and FT-IR spectroscopy. PGCMNPs were evaluated for in vitro antioxidant properties, uptake in HepG2 cells, biodistribution, in vivo toxicity and were also evaluated for anticancer therapeutic potential against NDEA-induced HCC in mice model in terms of tumor status, electrical properties, antioxidant defense status and apoptosis. The characterization studies confirmed successful formation of PGCMNPs with superparamagnetic properties. The internalization studies demonstrated (99-100)% uptake of PGCMNPs in HepG2 cells. These results were also supported by biodistribution studies in which increased iron content (296%) was noted inside the hepatocytes. Further, PGCMNPs exhibited no in vivo toxicity. The anticancer therapeutic potential was evident from observation that PGCMNPs treatment decreased tumor bearing animals (41.6%) and significantly (p ≤ 0.05) lowered tumor multiplicity. Overall, this study indicated that PGCMNPs with improved properties are efficiently taken-up by hepatoma cells and has therapeutic potential against HCC. Further, this agent can be tagged with 32P and hence can offer multimodal cancer treatment options via radiation ablation as well as magnetic hyperthermia.

Image dataset of Pune city historical places for degradation detection, classification, and restoration.

Historical and ancient murals hold clues to life in the past. This depicts the culture, worship styles, and social life of the community. This fascinates researchers and scholars who study historical bindings to showcase the modern world. Over time, most of these historical monuments have not been preserved in their original state. Most of these are affected due to natural climatic conditions, civil wars, and natural disasters. It remains clueless and imaginary about the size, shape, and color of the distorted historical monuments. This results in limitations for historical studies, archeological research, and geographical surveys. In this paper, we have studied the historical places around Pune city. Identified the locations where the monuments have been distorted and reconstructed recently. The construction age, type, color, shape, and size of the monuments are the major parameters of our study. Based on these criteria, we have captured images of these objects through different angles and camera lens. We have collected and categorized these images into folders with the names of historical places. This image dataset contains both captured and augmented images with distinct angles, scales, and directions. It also includes images captured in the daytime and evening with artificial lighting. This image dataset contains a variety of distinct image patterns that are useful as input to train computer-based supervised learning. The machine learning and deep learning algorithms perform efficiently if the input image dataset is large and distinct. Based on the predictive results generated by the machine learning and deep learning models, it is possible to virtually recreate the original monument. This would add a key value to historical research and studies.

Mito-TEMPO mitigates 5-fluorouracil-induced intestinal injury via attenuating mitochondrial oxidative stress, inflammation, and apoptosis: an in vivo study.

BACKGROUND: Recent evidences highlight role of mitochondria in the development of 5-fluorouracil (5-FU)-induced intestinal toxicity. Mitochondria-targeted antioxidants are well-known for their protective effects in mitochondrial oxidative stress- mediated diseases. In the present study, we investigated protective effect of Mito-TEMPO in 5-FU-induced intestinal toxicity. METHODS: Mito-TEMPO (0.1 mg/kg b.w.) was administered intraperitoneally to male BALB/c mice for 7 days, followed by co-administration of 5-FU for next 4 days (intraperitoneal 12 mg/kg b.w.). Protective effect of Mito-TEMPO on intestinal toxicity was assessed in terms of histopathological alterations, modulation in inflammatory markers, apoptotic cell death, expression of 8-OhDG, mitochondrial functional status and oxidative stress. RESULTS: 5-FU administered animals showed altered intestinal histoarchitecture wherein a shortening and atrophy of the villi was observed. The crypts were disorganized and inflammatory cell infiltration was noted. Mito-TEMPO pre-protected animals demonstrated improved histoarchitecture with normalization of villus height, better organized crypts and reduced inflammatory cell infiltration. The inflammatory markers and myeloperoxidase activity were normalized in mito-TEMPO protected group. A significant reduction in intestinal apoptotic cell death and expression of 8-OhDG was also observed in mito-TEMPO group as compared to 5-FU group. Further, mtROS, mtLPO and mitochondrial antioxidant defense status were improved by mito-TEMPO. CONCLUSION: Mito-TEMPO exerted significant protective effect against 5-FU-induced intestinal toxicity. Therefore, it may be used as an adjuvant in 5-FU chemotherapy.

Cardioprotective potential of mitochondria-targeted antioxidant, mito-TEMPO, in 5-fluorouracil-induced cardiotoxicity.

PURPOSE: The mitochondria-targeted antioxidants (MTAs) are known to offer protection against mitochondrial oxidative stress. The recent evidences support their role in mitigating oxidative stress-induced diseases, including cancer. Therefore, this study investigated cardioprotective potential of mito-TEMPO against 5-FU-induced cardiotoxicity. METHODS: Mito-TEMPO was administered to male BALB/C mice (intraperitoneally, 0.1 mg/kg b.w. for 7 days) followed by intraperitoneal administration of 5- FU (12 mg/kg b.w. for 4 days). During this period, mito-TEMPO treatment was also continued. The cardioprotective potential of mito-TEMPO was assessed by evaluating cardiac injury markers, extent of non-viable myocardium and histopathological alterations. Mitochondrial functional status and mitochondrial oxidative stress were assessed in cardiac tissue. 8-OHdG expression and apoptotic cell death were assessed using immunohistochemical techniques. RESULTS: The level of cardiac injury markers CK-MB and AST were significantly (P ≤ 0.05) decreased in mito-TEMPO pre-protected group which was further reflected in histopathology as decrease in the percentage of non-viable myocardial tissue, disorganization, and loss of myofibrils. Mito-TEMPO ameliorated mtROS, mtLPO and conserved mitochondrial membrane potential. Further, it had significantly (P ≤ 0.05) improved the activity of mitochondrial complexes and mitochondrial enzymes. A significant (P ≤ 0.05) increase in the level of mtGSH, activity of mitochondrial glutathione reductase, glutathione peroxidase, and mitochondrial superoxide dismutase was observed. A decreased expression of 8-OHdG and reduced apoptotic cell death were observed in mito-TEMPO pre-protected group. CONCLUSION: Mito-TEMPO effectively mitigated 5-FU-induced cardiotoxicity by modulating mitochondrial oxidative stress, hence may serve as a protective agent/adjuvant in 5-FU-based combinatorial chemotherapy.

Metformin and Dapagliflozin Attenuate Doxorubicin-Induced Acute Cardiotoxicity in Wistar Rats: An Electrocardiographic, Biochemical, and Histopathological Approach.

Doxorubicin is a widely used anticancer drug whose efficacy is limited due to its cardiotoxicity. There is no ideal cardioprotection available against doxorubicin-induced cardiotoxicity. This study aimed to investigate the anticipated cardioprotective potential of metformin and dapagliflozin against doxorubicin-induced acute cardiotoxicity in Wistar rats. At the beginning of the experiment, cardiac screening of experimental animals was done by recording an electrocardiogram (ECG) before allocating them into the groups. Thereafter, a total of thirty healthy adult Wistar rats (150-200 g) were randomly divided into five groups (n = 6) and treated for eight days as follows: group I (normal control), group II (doxorubicin control), group III (metformin 250 mg/kg/day), group IV (metformin 180 mg/kg/day), and group V (dapagliflozin 0.9 mg/kg/day). On the 7th day of the treatment phase, doxorubicin 20 mg/kg was administered intraperitoneal to groups II, III, IV, and V. On the 9th day (immediately after 48 h of doxorubicin administration), blood was collected from anesthetized animals for glucose, lipid profile, CK-MB & AST estimation, and ECG was recorded. Later, animals were sacrificed, and the heart was dissected for histopathological examination. We found that compared to normal control rats, CK-MB, AST, and glucose were significantly increased in doxorubicin control rats. There was a significant reversal of doxorubicin-induced hyperglycemia in the rats treated with metformin 250 mg/kg compared to doxorubicin control rats. Both metformin (180 mg/kg and 250 mg/kg) and dapagliflozin (0.9 mg/kg) significantly altered doxorubicin-induced ECG changes and reduced the levels of cardiac injury biomarkers CK-MB and AST compared to doxorubicin control rats. Metformin and dapagliflozin protected the cellular architecture of the myocardium from doxorubicin-induced myocardial injury. Current study revealed that both metformin and dapagliflozin at the FDA-recommended antidiabetic doses mitigated doxorubicin-induced acute cardiotoxicity in Wistar rats. The obtained data have opened the perspective to perform chronic studies and then to clinical studies to precisely consider metformin and dapagliflozin as potential chemoprotection in the combination of chemotherapy with doxorubicin to limit its cardiotoxicity, especially in patients with comorbid conditions like type II diabetes mellitus.

Phosphorylated chitosan accelerates dermal wound healing in diabetic wistar rats.

Phosphorylated chitosan (PC), a water-soluble derivative of chitosan possesses several biological and chemical properties suitable for diabetic wound healing. In the present study, we report the synthesis and diabetic wound healing capabilities of PC. Elemental analysis, FT-IR, 13C-NMR and 31P-NMR techniques were employed for the chemical characterization of PC. In vitro, antioxidant properties of PC were determined in terms of Fe3+ reducing, metal chelating, lipid peroxidation and superoxide scavenging ability. The wound healing potential of PC was assessed in diabetic excisional wound rat model. PC exhibited good water solubility, and in vitro antioxidant capacity. Wound contraction was higher in PC-treated wounds (91.11%) as compared to untreated wounds (67.26%) on 14th-day post wound creation. Histopathology of PC-treated wounds revealed improved tissue morphology with higher number of fibroblasts, a thicker epithelial layer, enhanced collagen deposits and angiogenesis as compared to untreated wounds. An overall increase of 57% and 25% in hydroxylamine and hexosamine content respectively were noted as compared to untreated wounds. A significant (P ≤ 0.05) increase in SOD activity and a significant (P ≤ 0.05) decrease in lipid peroxides were recorded in PC-treated wounds as compared to untreated wounds. These observations demonstrated that PC can be used as an effective agent in diabetic wound healing. Illustration of phosphorylated chitosan (PC) synthesis and its wound healing potential: Chitosan was phosphorylated to impart diabetic wound healing properties. Chemical characterizations such as elemental analysis, FT-IR and NMR confirmed successful phosphorylation of chitosan. PC exhibited good in vitro antioxidant properties. To assess the diabetic wound healing potential, an excisional wound model was developed in diabetic rats. PC treatment demonstrated accelerated wound healing.

Mito-TEMPO protects against bisphenol-A-induced testicular toxicity: an in vivo study.

Bisphenol-A (BPA) is a common environmental toxin which alters testicular function in both animals and humans. BPA exerts its cytotoxic potential by altering mitochondrial oxidative stress and functioning. Therefore, protecting mitochondria from oxidative stress may prevent BPA-induced testicular damage. In the present study, modulation of BPA toxicity by mitochondria-targeted antioxidant, mito-TEMPO was studied in male Wistar rats. Rats were administered mito-TEMPO (0.1 mg/kg b.w, i.p.) twice a week, followed by BPA (10 mg/kg b.w., orally) once a week for 4 weeks. After 4 weeks, sperm parameters were evaluated in the testis along with histopathological analysis. The mitochondrial oxidative stress, mitochondrial membrane potential (MMP) and enzymatic activity of mitochondrial complex II and IV were estimated in the testicular tissue. Pretreatment of mito-TEMPO protected animals from toxic effects of BPA as indicated by the normalization of sperm parameters and preserved histoarchitecture of testis. BPA treatment to animals significantly increased mitochondrial reactive oxygen species (ROS) and lipid peroxidation (LPO). A significant decrease in the activity of mitochondrial complex II was also observed after BPA exposure whereas, mitochondrial complex II activity was increased. In addition, an increase in MMP was also observed in BPA group. Mito-TEMPO successfully normalized mitochondrial ROS and LPO formation. Similar normalization effect was also noted in the activity of mitochondrial complex II, complex IV, and MMP. Results suggested that mito-TEMPO pretreatment significantly protected BPA-induced oxidative stress and thereby mito-TEMPO effectively prevented testicular damage.

Anticancer therapeutic potential of phosphorylated galactosylated chitosan against N-nitrosodiethyl amine-induced hepatocarcinogenesis.

Chitosan is a natural polyfunctional polymer that can be modified to achieve compounds with tailored properties for targeting and treating different cancers. In this study, we report the development and anticancer potential of phosphorylated galactosylated chitosan (PGC). The synthesized compound was characterized by FT-IR, NMR, and mass spectroscopy. The interaction of PGC with asialoglycoprotein receptors (ASGPR) and cellular internalization in HepG2 cells was studied using in silico and uptake studies respectively. PGC was evaluated for its metal chelating, ferric ion reducing, superoxide, and lipid peroxide (LPO) inhibiting potential. Further, anticancer therapeutic potential of PGC was evaluated against N-nitrosodiethylamine (NDEA)-induced hepatocellular carcinoma in a mice model. After development of cancer, PGC was administered to the treatment group (0.5 mg/kg bw, intravenously), once a week for 4 weeks. Characterization studies of PGC revealed successful phosphorylation and galactosylation of chitosan. A strong interaction of PGC with ASGP-receptors was predicted by computational studies and cellular internalization studies demonstrated 98.76 ± 0.53% uptake of PGC in the HepG2 cells. A good metal chelating, ferric ion reducing, and free radical scavenging activity was demonstrated by PGC. The anticancer therapeutic potential of PGC was evident from the observation that PGC treatment increased number of tumor free animals (50%) (6/12) and significantly (p ≤ 0.05) lowered tumor multiplicity as compared to untreated tumor group.

MitoQ demonstrates connexin- and p53-mediated cancer chemoprevention in N-nitrosodiethylamine-induced hepatocarcinogenesis rodent model.

Cancer chemoprevention is an approach that offers huge potential for preventing/retarding carcinogenesis. MitoQ is well-known and extensively studied mitochondria-targeted antioxidants for its applications in diseases linked with oxidative stress. In the present study chemopreventive potential of mitoQ was studied with a focus on the role of gap-junctions and p53 at an advanced stage of HCC. BALB/c mice model of hepatocarcinogenesis was established using N-nitrosodiethylamine as a carcinogen (200 mg/kg b. w., cumulative dose, intraperitoneally). The chemopreventive effect of mitoQ was studied by pre-protecting animals with mitoQ (0.125 mg/kg b. w., orally once a week) till the termination of the study. The tumors developed in the course of the study were histopathologically analyzed and statistically evaluated. The mechanistic role of mitoQ was investigated in terms of mitochondrial oxidative stress, expression of 8-OHdG, Cx26, Cx32, p53 and status of gap-junctional intercellular communication (GJIC) in tumors. Chemopreventive activity of mitoQ was evident from improved survival of animals, significantly (p ≤ 0.05) lower tumor multiplicity, tumor incidence and a total number of tumors. MitoQ treatment significantly (p ≤ 0.05) decreased mitochondrial oxidative stress as indicated by reduced mtROS and mtLPO. Increased staining intensity of 8-OHdG and internalization of Cx26, Cx32 which was observed in hepatic tumors was reduced upon mitoQ treatment. Furthermore, the expression of Cx26, Cx32 and p53 was significantly increased along with improvement in GJIC in mitoQ treatment group. MitoQ demonstrated its chemopreventive potential probably by regulating mtROS, connexins and p53 in hepatocarcinogenesis.

Quality Assessment of Tc-99m Methylene Diphosphonate (MDP) Radiopharmaceutical Prepared Using Different Cold Kit Fractionation Methods.

Purpose: Tc-99m Methylene diphosphonate (MDP) is prepared in house by labeling MDP cold kit. Each kit is a single time use vial and contains large amounts of reagent sufficient for preparing multiple doses. Therefore, several centers are adopting the practice of fractionating the MDP kit so that it can be used multiple times. The aim of the study was to evaluate the effect of kit fractionation on radiopharmaceutical property. Materials and Methods: The MDP kit was fractionated using two different approaches, namely, vial and syringe method. The quality of Tc-99m MDP prepared using these approaches was assessed and compared with that prepared by the conventional method. The image quality was evaluated in a total of 100 patients. Results: The vial and syringe fractionated Tc-99m MDP showed >95% RCP till the 4th and 2nd days of fractionation, respectively. Percentage radiochemical purity deteriorated to 83.6% and 88% on the 8th day of fractionation in the vial and syringe method, respectively. No microbial growth was observed in any of these methods till the 8th day of fractionation. The reconstituted MDP solution during all preparations was clear and colorless in appearance with pH ranging from 6.5 to 7.5. The image contrast, contrast-to-noise, and signal-to-noise ratio were statistically similar in both methods compared to the conventional method until the 2nd day of fractionation. The image quality data showed no statistical difference among images of vial and syringe fractionated MDP as compared to the conventional unfractionated Tc-99m MDP. Conclusions: The observations revealed that if fractionated with utmost care, both methods yield almost similar results.

Exploring New Drug Targets for Type 2 Diabetes: Success, Challenges and Opportunities.

There are substantial shortcomings in the drugs currently available for treatment of type 2 diabetes mellitus. The global diabetic crisis has not abated despite the introduction of new types of drugs and targets. Persistent unaddressed patient needs remain a significant factor in the quest for new leads in routine studies. Drug discovery methods in this area have followed developments in the market, contributing to a recent rise in the number of molecules. Nevertheless, troubling developments and fresh challenges are still evident. Recently, metformin, the most widely used first-line drug for diabetes, was found to contain a carcinogenic contaminant known as N-nitroso dimethylamine (NDMA). Therefore, purity and toxicity are also a big challenge for drug discovery and development. Moreover, newer drug classes against SGLT-2 illustrate both progress and difficulties. The same was true previously in the case of glucagon-like peptide-1 receptor agonists and dipeptidyl peptidase-4 inhibitors. Furthermore, researchers must study the importance of mechanistic characteristics of novel compounds, as well as exposure-related hazardous aspects of current and newly identified protein targets, in order to identify new pharmacological molecules with improved selectivity and specificity.

Adjunctive Diagnostic Methods for Skin Cancer Detection: A Review of Electrical Impedance-Based Techniques.

Skin cancer is among the fastest-growing cancers with an excellent prognosis, if detected early. However, the current method of diagnosis by visual inspection has several disadvantages such as overlapping tumor characteristics, subjectivity, low sensitivity, and specificity. Hence, several adjunctive diagnostic techniques such as thermal imaging, optical imaging, ultrasonography, tape stripping methods, and electrical impedance imaging are employed along with visual inspection to improve the diagnosis. Electrical impedance-based skin cancer detection depends upon the variations in electrical impedance characteristics of the transformed cells. The information provided by this technique is fundamentally different from other adjunctive techniques and thus has good prospects. Depending on the stage, type, and location of skin cancer, various impedance-based devices have been developed. These devices when used as an adjunct to visual methods have increased the sensitivity and specificity of skin cancer detection up to 100% and 87%, respectively, thus demonstrating their potential to minimize unnecessary biopsies. In this review, the authors track the advancements and progress made in this technique for the detection of skin cancer, focusing mainly on the advantages and limitations in the clinical setting. © 2022 Bioelectromagnetics Society.

Optical Trapping and Micro-Raman Spectroscopy of Functional Red Blood Cells Using Vortex Beam for Cell Membrane Studies.

There has been a long-standing interest in Raman spectroscopic investigation of optically trapped single functional cells. Optical trapping using a Gaussian beam has helped researchers for decades to investigate single cells suspended in a physiological medium. However, complete and sensitive probing of single cells demands further advancements in experimental methods. Herein, we propose optical trapping and simultaneous micro-Raman spectroscopy of red blood cells (RBCs) in an unconventional face-on orientation using an optical vortex beam. Using this novel method, we are successful in comparing the conformational state of hemoglobin (Hb) molecules near the RBC membrane and inside the bulk of the cell. This method enabled us to successfully probe the oxy/deoxy ratio of Hb molecules near the RBC membrane and inside the bulk of the cell. Because of the face-on orientation, the Raman spectra of RBCs acquired using a vortex beam have a significant contribution from membrane components compared to that recorded using the Gaussian beam.

Mitochondria-targeted antioxidant, mito-TEMPO mitigates initiation phase of N-Nitrosodiethylamine-induced hepatocarcinogenesis.

Targeting mitochondrial oxidative stress during initial stages of hepatocarcinogenesis can be an effective and promising strategy to prevent hepatocellular carcinoma (HCC). In the present study, mitochondria targeted antioxidant, mito-TEMPO was administered to male BALB/c mice at a dosage 0.1 mg/kg b.w. (intraperitoneal) twice a week, followed by single N-Nitrosodiethylamine (NDEA) intraperitoneal injection (10 mg/kg b.w.). After 24 h of NDEA administration, animals were sacrificed, blood and liver tissue were collected. Liver injury markers, histoarchitecture, antioxidant defence status, mitochondrial reactive oxygen species (ROS), lipid peroxidation (LPO), mitochondrial dysfunction analysis, and mitochondrial membrane potential were investigated. Mito-TEMPO pre-treatment protected animals from the damaging effects of NDEA as observed by normalization of liver injury markers. NDEA metabolism resulted in a significantly increased intracellular and mitochondrial ROS generation with concomitant increase in LPO formation. The activity of mitochondrial complex I, complex II, malate dehydrogenase were significantly reduced and mitochondrial membrane potential was increased. Mito-TEMPO effectively scavenged NDEA-induced ROS generation and reduced LPO formation. A significant improvement was also observed in the activity of mitochondrial complex I, complex II, malate dehydrogenase and normalisation of mitochondrial membrane potential. Results suggested that mito-TEMPO had significant impact on the initiation phase of hepatocarcinogensis which could be one of the reason for its reported chemopreventive effect.

Selenium attenuates venlafaxine hydrochloride-induced testicular damage in mice via modulating oxidative stress and apoptosis.

The present study assessed the effect of selenium (Se) supplementation on Venlafaxine hydrochloride (VH)-induced testicular toxicity. Mice were segregated into Group I (C), Group II (0.5 ppm Se), Group III (VH at a dose 60 mg/kg b.w.) and Group IV (Se was given as per Group II, and VH was given as per Group III). After 10 weeks, sperm parameters, histology, sperm cell counts, antioxidants activities, apoptotic proteins and molecular analysis of testicular tissue were evaluated. Group III had significantly lower sperm concentration (from 2.17 ± 0.28 to 1.04 ± 0.22) and sperm motility (from 68.04 ± 5.5 to 21.47 ± 5.21), and showed an extensive vacuolisation in the germinal epithelium, abnormal basement membrane, and reduced germ cell number as compared to Group I. However, selenium supplementation in Group IV substantially increased sperm concentration (1.47 ± 0.48) and motility (33.27 ± 8.66), improved the histoarchitecture and repopulated the germ cells as observed by raised numbers of spermatogonia, spermatocytes, round spermatids and elongated spermatids contrasted to Group III. Group IV also showed a noteworthy decreased ROS, LPO levels, as well as expressions of Bax, caspase-9, and caspase-3 and increased the SOD, CAT, GPx, and GSH activities as well the expression of Bcl-2 as compared to Group III. This effect was further supported by FTIR analysis for nucleic acids. Thus, selenium supplementation showed significant protection against VH-induced testicular toxicity.

Electrical conductivity spectra of hepatic tumors reflect hepatocellular carcinoma progression in mice.

Background:Electrical impedance spectroscopy is a technique which evaluates differences in dielectric properties of tissues for cancer identification.Methods:Murine hepatic cancer model was developed by intraperitoneal administration of N-nitrosodiethylamine to male BALB/c mice. Tumors obtained were evaluated for their conductivity in frequency range of (4 Hz-5 MHz). All tumors were subjected to histopathological grading and parameters such as free spacing, necrosis, and cell density were estimated on histological slides. The status of gap junctions and gap junction intercellular communication (GJIC) were studied using enzyme-linked immunosorbent assay, immunohistochemistry, dye transfer assay, and electron microscopy.Results:Histopathological investigation revealed the presence of moderately to poorly-differentiated hepatocellular carcinoma (HCC) in mice. All types of tumors showed higher electrical conductivity than normal liver tissue in frequency range (4 Hz-1 kHz). However, in frequency range (10 kHz-5 MHz) only poorly-differentiated tumors showed higher conductivity compared to normal tissue. The most prominent findings in moderately-differentiated and poorly-differentiated HCC were increased visible free spaces and necrosis respectively. The status of cell gap junctions were significantly deteriorated in tumors and a corresponding significant reduction in GJIC was also observed. These biological indicators were correlated with electrical conductivity of hepatic tumors.Conclusion:Variations in electrical conductivity spectra of hepatic tumors reflect progression of HCC.General significance:Future studies can be planned to perform hierarchical clustering of dielectric parameters with more number of tumor samples to establish dielectric spectroscopy-based classification or staging of hepatic tumors.

Mito-TEMPO, a mitochondria-targeted antioxidant, prevents N-nitrosodiethylamine-induced hepatocarcinogenesis in mice.

BACKGROUND: Oxidative stress and mitochondrial dysfunction play a significant role in hepatocarcinogenesis. Mitochondria are source organelle as well as target for free radicals. The oxidative damage to mitochondria can be prevented by mitochondria-targeted antioxidant, mito-TEMPO. However, its efficacy in prevention of hepatocellular carcinoma has not been investigated so far. METHODS: Murine model of hepatocarcinogenesis was developed by intraperitoneal administration of N-nitrosodiethylamine to male BALB/c mice. Mito-TEMPO was administered intraperitoneally at weekly intervals, till the completion of the study. The tumours were histopathologically analysed and anti-cancer efficacy of mito-TEMPO was evaluated in terms of survival index, tumour incidence, tumour multiplicity and tumour dielectric parameters. The antioxidant defence status and molecular composition of tumours were assessed. Gap junctions and gap-junctional intercellular communication (GJIC) were studied using ELISA, IHC and Lucifer yellow assay. RESULTS: Mito-TEMPO treatment increased survival of animals by 30%, decreased tumour incidence (25%) and tumour multiplicity (39%). The dielectric parameters of tumours in Mito-TEMPO group were indicative of retarded carcinogenesis. Mito-TEMPO administration normalized mean saturation levels in phospholipids and improved glycogen content of the hepatic tissue. Gap junctions and GJIC which were severely impaired in hepatocarcinogenesis, improved after mito-TEMPO treatment. CONCLUSION: Mito-TEMPO was effective in combating hepatocarcinogenesis.

Peptides as adjuvants for ampicillin and oxacillin against methicillin-resistant Staphylococcus aureus (MRSA).

Fast emerging antibiotic resistance in pathogens requires special attention for strengthening the reservoir of antimicrobial compounds. In view of this, several peptides with known antimicrobial activities have been reported to enhance the efficacy of antibiotics against multidrug resistant (MDR) pathogens. In the present study, potential of peptides having distinct mechanism of action, if any, was evaluated to improve the efficacy of conventional antibiotics against methicillin-resistant S. aureus (MRSA). After primary screening of six peptides, two peptides namely T3 and T4 showing very high minimum inhibitory concentrations (MICs) were selected to assess their role in altering the MICs of antibiotics to which the pathogen was resistant. In the presence of the peptides, the MICs of the antibiotics were found to be reduced as per the fractional inhibitory concentration indices (FICI) and time kill assay. These observations prompted us to look for their mechanism of action. The effect of peptides on the morphology of pathogen by field emission scanning electron microscopy (FE-SEM) revealed no damage to the cells at the sub-inhibitory concentrations of the peptide which correlated well with the higher MIC of the peptide, indicating no direct impact on the pathogen. However, dielectric spectroscopy, confocal microscopy and flow cytometry confirmed the interaction and localization of peptides with the bacterial membrane. The peptides were also found to inhibit efflux of ethidium bromide which is the substrate for many proteins involved in efflux system. Therefore, it is speculated that the peptides after interacting with the membrane of the pathogen might have resulted in the inhibition of the efflux of antibiotics thereby reducing their effective concentrations. The study thus suggests that peptides with no antimicrobial activity of their own, can also enhance the efficacy of the antibiotics by interacting with the pathogen thereby, acting as adjuvants for the antibiotics.

Preventive Role of Vitamin D in Silica-Induced Skin Fibrosis: A Study in Relation to Oxidative Stress and Pro-Inflammatory Cytokines.

The protective effects of vitamin D analogue calcipotriol in silica-induced skin fibrosis were studied in the present study. Male BALB / c mice were divided into four groups; Control, Vitamin D, Silica and Silica+Vitamin D. Silica was administered as a single intradermal injection (40 µg / µL, dissolved in normal saline; particle size 1 - 5 µm) in the hind limbs of animals in Silica & Silica+Vitamin D group. Vitamin D group animals received topical application of 100µL of vitamin D solution (10-7M in Ethanol) daily for 12 weeks. Silica+Vitamin D group animals received co-treatment of silica and vitamin D as described for other groups. After 12 weeks of treatment, dermal thickness and hydroxyproline content of treated sections were measured. The TNF-α and IL-6 levels were measured in serum of all treated animals. The silica-induced oxidative stress was measured in terms of lipid peroxidation in skin tissue. Antioxidant defence system was assessed in terms of levels of reduced glutathione, superoxide dismutase, catalase, glutathione peroxidase and glutathione reductase. A significant increase in the dermal thickness and hydroxyproline content was observed after silica treatment (931 ± 57.98 to 1804.61 ± 146.20 µm)(p < 0.05). Vitamin D co-treatment reduced dermal thickness and hydroxyproline content compared to Silica group (p < 0.05). Similarly a decrease in TNF-α and IL-6 levels were also observed after vitamin D treatment. A significant reduction in oxidative stress in terms of lipid peroxidation (4.92 ± 0.70 to 2.40 ± 0.31 nmol / mg protein). Therefore, present study suggested that vitamin D could be an effective agent against silica-induced skin fibrosis and oxidative stress.

Fourier Transform Infrared Spectroscopic Studies on Modulation of N-Nitrosodiethylamine-Induced Hepatocarcinogenesis by Azadirachta indica.

Fourier transform infrared spectroscopy was employed in the present study to obtain information about the molecular composition of hepatic tumor versus hepatic tissue. A hepatic cancer model was developed by administering N-nitrosodiethylamine (NDEA) to male Balb/c mice. The results revealed that NDEA-induced hepatic cancer tumor tissue had altered molecular composition compared with normal liver tissue. Compared with the normal tissue, the saturation level of membrane phospholipids was observed to be decreased in tumors along with an abnormal distribution of protein secondary structures. A significant decrease in glycogen and a significant increase in total nucleic acid content were also observed in tumor cells. The administration of aqueous Azadirachta indica leaf extract (AAILE) prior to NDEA treatment resulted in the normalization of saturation levels in phospholipids and total nucleic acid content and in the distribution of protein secondary structures in tumors. A significant increase in the amount of stored glycogen was observed in AAILE cotreated tumors compared with NDEA-induced tumors, which might indicate that AAILE cotreatment impeded the ability of tumor cells to consume glucose at a faster rate. The normalization of molecular composition upon AAILE cotreatment in hepatic tumors might indicate that AAILE hampered the process of evolution of tumors, which could be responsible for its observed chemopreventive action.