Essential oil of Eucalyptus citriodora: Physio-Chemical analysis, Formulation with Hand Sanitizer Gel and Antibacterial Activity

Background: The essential oil-bearing plants are extensively being used in traditional systems of medicine due to the occurrence of the diversity of phytochemical constituents. The emerging crisis of developing resistance to conventional drugs has increased public health awareness and reliance on natural compounds as safer alternatives.Methods: The essential oil extracted from Eucalyptus citriodora (Hook.) leaves was characterized for physicochemical attributes, formulated with hand sanitizer gel, tested for organoleptic parameters, and antimicrobial activity against Escherichia coli, Staphylococcus aureus, and Bacillus subtilis.Results: E. citriodora essential oil (EEO) had a camphorous scent, and dark yellow coloration, while exhibiting 0.60% yield (v/w, 97% pure), 0.94 density, 1.47 refractive index, 11.10 viscosity, 0.92 specific gravity, 0.0-9.98º optical rotation, 11.20 acid number, 50.60 ester number, which satisfy the standards specified by ISO (The International Organization for Standardization). The chromatographic analysis of oil identified eucalyptol as the most abundant compound (80.08%) followed by α-terpinyl acetate, isopinocarveol, and globulol as the moderately abundant compounds (4.46-4.81%), while viridiflorol and terpinen-4-ol as less abundant compounds (3.06 and 2.69%, respectively). Formulated hand sanitizer with EEO exhibited physical and microbiological properties that were comparable with the market products. It also had a pleasant scent, was compatible with the skin, was easy to apply, and is acceptable to the users.Conclusion: The current study clearly shows that EEO could be utilized as a potential ingredient in alcohol-based gel hand sanitizer formulation for giving a pleasant smell, acceptable physical appearance and microbial quality parameters.Keywords: Essential oil; Eucalyptol; Hand sanitizer; Gel Formulation; Antimicrobial effect

Cucurbitacin D ameliorates benzo[a]pyrene induced liver injury via Activation of Nrf2 Antioxidant pathway

Background: Co-morbidity variables, such as smoking, are strongly linked to the development and progression of liver cancer. Further, benzo[a]pyrene, a major component of tobacco smoke, is highly carcinogenic and triggers liver damage. Cucurbitacin, a kind of triterpene, has a wide range of biological properties, including antioxidant, anti-inflammatory, and anti-cancer effects. However, the precise mechanism of its hepatoprotective effects is obscure. Objective: The aim of this study is to investigate the cytoprotective effects of novel analog of cucurbitacin, cucurbitacin D, against benzo[a]pyrene-induced liver injury in HepG2 cells. Method: The cytoprotective efficacy of cucurbitacin D against benzo[a]pyrene-induced liver damage was studied using proliferation, clonogenicity, migration, invasion, Western blotting, and qPCR analysis. The levels of intracellular reactive oxygen species (ROS) in liver cells was measured using the DCFDA assay. Results: In human HepG2 cells, functional experiments revealed that cucurbitacin D has cytoprotective effects against dose-dependent growth inhibition by benzo[a]pyrene. The mitigation of ROS observed by fluorimeter and fluorescence microscopy suggested that this protective effect was likely due to cucurbitacin D\u27s antioxidant property. Additional research is ongoing to identify the effect of cucurbitacin D on oxidative stress markers by using qPCR and western blotting techniques. Overall, these findings showed that cucurbitacin D diminishes benzo[a]pyrene-induced liver injury via its antioxidant activity. Conclusion: These findings show that cucurbitacin D has hepatoprotective properties against benzo[a]pyrene-induced liver injury, making it an attractive food supplement ingredient

Use of Endoscopic Third Ventriculostomy (ETV) Success Score to Predict the Outcome of Endoscopic Third Ventriculostomy in Obstructive Hydrocephalus

Object: In February 2011 Abhaya V. Kulkarni et al. reported endoscopic third ventriculostomy success score in order to predict the outcome of obstructive hydrocephalus. After ETV the object of the present study was to evaluate this predictive value in our own setup, prospectively.Materials and Methods: From April 2011-November 2012, 110 endoscopic third ventriculostomy procedures were performed for obstructive hydrocephalus of different etiologies at the department of pediatric neurosurgery, the Children’s Hospital Lahore. All of these cases were more than 6 months old and the data was analyzed by the senior author.Results: A total number of 110 patients were operated between April 2011 – November 2012. The patients’ ages ranged from 6 months – 13 years. No child below 6 months was included in this study. Children were stratified into 4 age groups: 6 months – 1 year (Group – 1), 1 year – 2 years (Group – 2), 2 years – 10 years (Group – 3) and more than 10 years (Group – 4). The score was calculated for each patient before surgery according to ETVSS and at the end of 6months the success or failure of the ETV was determined by clinical, radiological measures. Out of 110 patients, only 80 were available at the completion of 6months period after surgery. The ETV was successful in 56 patients (70%). Patients below 1 year achieved lowest success. Of the ten patients with a high probability of ETV success, eight (80%) were successfully treated.Conclusion: The results show that EVT success can be predicted very well by ETVSS and it should help in establishing surgical selection criteria in order to obtain high success rate. It can also help in preparing the patients and their families to the expected outcome

Hepatoprotective role of Cucurbitacin D on benzo[a]pyrene induced liver injury

Background: Epidemiological findings show the strong correlation of co-morbidity factors including smoking with the development and progression of liver cancer. Moreover, benzo[a]pyrene, a main component of tobacco smoke, is extremely carcinogenic and contributes to liver injury as well. Cucurbitacin, chemically classified as triterpenes, have shown diverse biological activities including potent antioxidant, anti-inflammatory and anti-cancer activities. However, their hepatoprotective activities are not completely understood. Objective: In the present study, we investigated the cytoprotective activity of novel analog of cucurbitacin, cucurbitacin D, against benzo[a]pyrene-induced liver injury in human HepG2 cells. Method: Proliferation, clonogenicity, migration, invasion, Western blotting and qPCR analysis were conducted to investigate the cytoprotective effect of cucurbitacin D against benzo[a]pyrene induced liver damage. DCFDA assay was performed to analyze intracellular reactive oxygen species (ROS) level in liver cells. Results: Functional assays showed that cucurbitacin D exhibited cytoprotective effects against dose-dependent growth inhibition by benzo[a]pyrene in human HepG2 cells. This protective effect was likely associated with antioxidant potential of cucurbitacin D, as evidenced by the attenuation of ROS observed by fluorimeter and fluorescence microscopy. Further study is ongoing to examine the effect of cucurbitacin D on oxidative stress markers by employing western blotting and qPCR techniques. Collectively, these results exhibited that cucurbitacin D alleviate benzo[a]pyreneinduced liver injury through its antioxidant effects. Conclusion: These results have demonstrated hepatoprotective effects of cucurbitacin D against benzo[a]pyrene-induced liver damage, rendering it as an effective potential ingredient in food supplements

Therapeutic efficacy of ormeloxifene against hepatocellular carcinoma

Background: Hepatocellular carcinoma (HCC) is the most common primary liver cancer and leading cause of cancer related deaths worldwide. Severe toxicity and drug resistance to available chemotherapeutic agents display ineffective clinical response. Therefore, drug repurposing is gaining attention owing to their known biological activities and excellent safety profiles. Ormeloxifene (ORM), non-steroidal, selective estrogen receptor modulator (SERM), and exhibit diverse pharmacological activities. The aim of this study is to assess the therapeutic activity of ORM and to investigate the underlying molecular mechanism against hepatocellular carcinoma. Objective: To investigate the therapeutic activity of ormeloxifene in human hepatocellular carcinoma cells. Methodology: MTT and colony formation assays were performed in SK-Hep-1, Hep3B and C3A cells. In vitro functional assays were carried out for investigating effect of ORM on migration and invasion abilities of HCC cells using Boyden chamber and Matrigel assays respectively. Results: Functional analysis revealed that ORM treatment led to suppression of proliferation and colony formation in human hepatocellular carcinoma cells in dose and time-dependent manner compared to vehicle treated group. ORM treatment, as shown by wound healing and Matrigel invasion assay, respectively, suppresses the migration and invasion of human hepatocellular carcinoma cells. Further, experiments are underway to determine the effect of ORM on EMT markers using western blotting and qPCR techniques. Conclusion: Taken together, ORM exhibited potent anticancer effects against HCC and could be further explored as a novel therapeutic modality for the treatment of HCC

Pharmacological restoration of PKD1: A novel strategy for prostate cancer therapy

Background: Prostate cancer has poor prognosis owing to late diagnosis and ineffective multimodal clinical treatment. Extensive efforts are ongoing to establish methods that can resolve the expression of genes implicated in disease development and treatment. Previously, we reported that Protein Kinase D1 (PKD1), a serine threonine kinase, controls a number of tumor suppressor functions including cell aggregation, cell motility, cell proliferation, and cell invasion. Thus, PKD1 is considered as an emerging therapeutic target for prostate cancer treatment. Objective: To investigate the restoration of PKD1 by a pharmacological modulator ormeloxifene, which showed well-defined PK/PD and safety profiles in humans. Methods: Proliferation, clonogenicity, migration, invasion, western blotting and qPCR analysis were performed to investigate the anticancer effect of ORM, docetaxel and/or their combination on PKD1 and related signaling mechanisms in prostate cancer. Results: ORM treatment inhibited cell proliferation, invasion, migration and colony formation abilities of prostate cancer cells in a dose-dependent manner compared to vehicle treated group. ORM treatment selectively induces the expression of PKD1 both at mRNA and protein levels in C4-2 cells. Moreover, our results have also shown that ORM effectively attenuates MTA1 expression in prostate cancer cells. MTA1 physically interact and shown to have inverse relationship with PKD1. In addition, we observed that ORM treatment enhances the therapeutic efficacy of docetaxel in C4-2 cells. Our results also indicate that ORM treatment potentiate the effects of docetaxel as determined by MTS and colony formation assays. Conclusion: These results suggest that ORM exhibit potent anticancer activity via restoration of PKD1 in prostate cancer

Mucin 13 expression correlates with tumor development in hepatocellular carcinoma

Background: Hepatocellular carcinoma (HCC) has a poor prognosis due to ineffective therapeutic modality and lack of early diagnostic marker. Accumulating studies have shown that elevated expression of mucin 13 as potential oncogene and predictive biomarker for various cancer. However, very little is known about its expression and function for development and progression of HCC Objective: To investigate mucin 13 expression in chemically induced hepatocellular carcinoma model. Methodology: Diethyl nitrosamine (DEN) and 2-Acetylaminofluorene (2-AAF) induced method was employed for the development of hepatocellular carcinoma in Male Wistar rats. Serum and tissues were collected at regular interval of time and routinely validated for liver cancer stages. Immunohistochemistry and in situ hybridization were performed on formalin-fixed, paraffin-embedded tissues. Molecular docking studies were performed to study the interaction of mucin 13 and DEN. Results: Our results demonstrate hepatocellular adenoma as observed by histopathological analysis. Biochemical analysis showed a progressive increase in the levels of serum ALT, AST and ALP, suggesting the development and progression of hepatocellular damage. Notably, mucin 13 expression gradually elevated during consecutive stages of hepatocellular carcinoma. Interestingly, an increase in nuclear localization of mucin 13 was observed in treated group as compared to control group. In situ hybridization analysis showed that a decrease in miR-132 and miR-145, which are inversely related with mucin 13 expression. Moreover, DEN efficiently binds mucin 13 with high affinity and thus stabilize it as demonstrated by molecular docking analysis. Conclusion: These results suggest that mucin 13 expression is closely associated with hepatocarcinogenesis and could serve as a predictive candidate biomarker for HCC

Molecular Insights into Targeting PKD1 for Prostate Cancer Treatment

Background: Prostate cancer has a poor prognosis due to late diagnosis and ineffective multimodal clinical treatment. Efforts are underway to create strategies for resolving the abnormal expression of molecular targets implicated in disease development and progression. We previously reported that the serine threonine kinase Protein Kinase D1 (PKD1) regulates a multitude of tumor suppressor functions, including cell aggregation, motility, proliferation, and invasion in prostate cancer. Thus, PKD1 is regarded as a promising therapeutic target for the treatment of prostate cancer. Objective: The goal of this study was to investigate the therapeutic potential of ormeloxifene (ORM), a pharmacological modulator with well-defined PK/PD and safety profiles in humans, for PKD1 restoration in prostate cancer. Methods: The anticancer effect of ORM on PKD1 and associated signaling mechanisms in prostate cancer was investigated using proliferation, clonogenicity, migration, invasion, western blotting, and qPCR analysis. Results: In comparison to the vehicle-treated group, ORM treatment decreased prostate cancer cell proliferation, invasion, migration, and colony formation in a dose-dependent manner. In C4-2 cells, ORM treatment selectively induces PKD1 expression at both the mRNA and protein levels. Furthermore, our findings revealed that ORM efficiently suppresses MTA1 expression in prostate cancer cells. MTA1 physically interacts with PKD1 and has been shown to have an inverse correlation with it. Our results also showed that ORM treatment enhances the therapeutic efficacy of decetaxel. Conclusion: Taken together, these findings show that ORM has anticancer properties in prostate cancer via restoring PKD1

Mucin 13 expression is an early indicator of hepatocellular carcinoma development

Background: Hepatocellular carcinoma (HCC) has a poor prognosis due to ineffective therapeutic modality and lack of an early marker for diagnosis. Studies show that increased mucin 13 (MUC13) expression as a possible oncogene and predictive biomarker for various cancers has been shown. But its expression and role in the development of HCC is very little known. Objective: The aim of this study is to investigate the MUC13 expression in chemically induced hepatocellular carcinoma model. Methodology: Male Wistar rats were subjected to a DEN and 2-Acetylaminofluorene (2-AAF) induced method for the development of hepatocellular carcinoma. Serum and tissues were collected at regular intervals and routinely validated for various stages of liver cancer. On formalin-fixed, paraffin-embedded tissues, immunohistochemistry and in situ hybridization were performed. The molecular interaction of mucin 13 and DEN were also performed using in silico analysis. Results: Histopathological analysis of liver tissues revealed the development of hepatocellular carcinoma with successive stages in chemically induced model HCC. Moreover, biochemical analysis showed a progressive increase in serum ALT, AST, and ALP levels, indicating the development and progression of hepatocellular damage. Notably, mucin 13 expression gradually increased during the progression of hepatocellular carcinoma. The treated group showed an increase in nuclear localization of mucin 13 as compared to the control group. In situ hybridization analysis revealed a reduction in miR-132 and miR145, both of which are inversely related to mucin 13 expression. Furthermore, molecular docking analysis showed that DEN efficiently binds mucin 13 with high affinity and thus stabilizes it. Conclusion: These findings suggest that mucin 13 expression is linked to hepatocarcinogenesis and could be used as a candidate biomarker for HCC

Cucurbitacin D exhibits potent anticancer activity in cervical cancer

In this study, we for the first time, investigated the potential anti-cancer effects of a novel analogue of cucurbitacin (Cucurbitacin D) against cervical cancer in vitro and in vivo. Cucurbitacin D inhibited viability and growth of cervical cancer cells (CaSki and SiHa) in a dose-dependent manner. IC50 of Cucurbitacin D was recorded at 400 nM and 250 nM in CaSki and SiHa cells, respectively. Induction of apoptosis was observed in Cucurbitacin D treated cervical cancer cells as measured by enhanced Annexin V staining and cleavage in PARP protein. Cucurbitacin D treatment of cervical cancer cells arrested the cell cycle in G1/S phase, inhibited constitutive expression of E6, Cyclin D1, CDK4, pRb, and Rb and induced the protein levels of p21 and p27. Cucurbitacin D also inhibited phosphorylation of STAT3 at Ser727 and Tyr705 residues as well as its downstream target genes c-Myc, and MMP9. Cucurbitacin D enhanced the expression of tumor suppressor microRNAs (miR-145, miRNA-143, and miRNA34a) in cervical cancer cells. Cucurbitacin D treatment (1 mg/kg body weight) effectively inhibited growth of cervical cancer cells derived orthotopic xenograft tumors in athymic nude mice. These results demonstrate the potential therapeutic efficacy of Cucurbitacin D against cervical cancer