Molecular Targets of Plant-based Alkaloids and Polyphenolics in Liver and Breast Cancer- An Insight into Anticancer Drug Development.

Liver and Breast cancer are ranked as the most prevailing cancers that cause high cancer-related mortality. As cancer is a life-threatening disease that affects the human population globally, there is a need to develop novel therapies. Among the available treatment options include radiotherapy, chemotherapy, surgery, and immunotherapy. The most superlative modern method is the use of plant-derived anticancer drugs that target the cancerous cells and inhibit their proliferation. Plant-derived compounds are generally considered safer than synthetic drugs/traditional therapies and could serve as potential novel targets to treat liver and breast cancer to revolutionize cancer treatment. Alkaloids and Polyphenols have been shown to act as anticancer agents through molecular approaches. They disrupt various cellular mechanisms, inhibit the production of cyclins and CDKs to arrest the cell cycle, and activate the DNA repairing mechanism by upregulating p53, p21, and p38 expression. In severe cases, when no repair is possible, they induce apoptosis in liver and breast cancer cells by activating caspase-3, 8, and 9 and increasing the Bax/Bcl-2 ratio. They also deactivate several signaling pathways, such as PI3K/AKT/mTOR, STAT3, NF-kB, Shh, MAPK/ERK, and Wnt/ß-catenin pathways, to control cancer cell progression and metastasis. The highlights of this review are the regulation of specific protein expressions that are crucial in cancer, such as in HER2 over-expressing breast cancer cells; alkaloids and polyphenols have been reported to reduce HER2 as well as MMP expression. This study reviewed more than 40 of the plant-based alkaloids and polyphenols with specific molecular targets against liver and breast cancer. Among them, Oxymatrine, Hirsutine, Piperine, Solamargine, and Brucine are currently under clinical trials by qualifying as potent anticancer agents due to lesser side effects. As a lot of research is there on anticancer compounds, there is a desideratum to compile data to move towards clinical trials phase 4 and control the prevalence of liver and breast cancer.

Addressing artifacts of colorimetric anticancer assays for plant-based drug development.

Cancer has become the silent killer in less-developed countries and the most significant cause of morbidity worldwide. The accessible and frequently used treatments include surgery, radiotherapy, chemotherapy, and immunotherapy. Chemotherapeutic drugs traditionally involve using plant-based medications either in the form of isolated compounds or as scaffolds for synthetic drugs. To launch a drug in the market, it has to pass through several intricate steps. The multidrug resistance in cancers calls for novel drug discovery and development. Every year anticancer potential of several plant-based compounds and extracts is reported but only a few advances to clinical trials. The false-positive or negative results impact the progress of the cell-based anticancer assays. There are several cell-based assays but the widely used include MTT, MTS, and XTT. In this article, we have discussed various pitfalls and workable solutions.

Synthesis of Silver Nanoparticles from Extracts of Wild Ginger (Zingiber zerumbet) with Antibacterial Activity against Selective Multidrug Resistant Oral Bacteria.

Antibiotic resistance rate is rising worldwide. Silver nanoparticles (AgNPs) are potent for fighting antimicrobial resistance (AMR), independently or synergistically. The purpose of this study was to prepare AgNPs using wild ginger extracts and to evaluate the antibacterial efficacy of these AgNPs against multidrug-resistant (MDR) Staphylococcus aureus, Streptococcus mutans, and Enterococcus faecalis. AgNPs were synthesized using wild ginger extracts at room temperature through different parameters for optimization, i.e., pH and variable molar concentration. Synthesis of AgNPs was confirmed by UV/visible spectroscopy and further characterized by scanning electron microscopy (SEM), X-ray diffraction (XRD), energy-dispersive X-ray spectroscopy analysis (EDXA), and Fourier-transform infrared spectroscopy (FTIR). Disc and agar well diffusion techniques were utilized to determine the in vitro antibacterial activity of plant extracts and AgNPs. The surface plasmon resonance peaks in absorption spectra for silver suspension showed the absorption maxima in the range of 400-420 nm. Functional biomolecules such as N-H, C-H, O-H, C-O, and C-O-C were present in Zingiber zerumbet (Z. zerumbet) (aqueous and organic extracts) responsible for the AgNP formation characterized by FTIR. The crystalline structure of ZZAE-AgCl-NPs and ZZEE-AgCl-NPs was displayed in the XRD analysis. SEM analysis revealed the surface morphology. The EDXA analysis also confirmed the element of silver. It was revealed that AgNPs were seemingly spherical in morphology. The biosynthesized AgNPs exhibited complete antibacterial activity against the tested MDR bacterial strains. This study indicates that AgNPs of wild ginger extracts exhibit potent antibacterial activity against MDR bacterial strains.

Anal human papillomavirus infection among men who have sex with men and transgender women living with and without HIV in Pakistan: findings from a cross-sectional study.

OBJECTIVES: The aim of this study was to determine the prevalence of infection, genotypes and risk factors for human papillomavirus (HPV) among men who have sex with men (MSM) and transgender women living with and without HIV in Pakistan. Anal infection with HPV is very common worldwide among MSM, particularly among MSM living with HIV. The high prevalence of HIV among MSM and male-to-female transgendered individuals in Pakistan is a significant health concern since access to screening and health-seeking is often delayed in this stigmatised key population. DESIGN: This cross-sectional study was conducted between March 2016 and November 2017. PARTICIPANTS, SETTING AND DATA COLLECTION: This study recruited MSM and transgender-women who self-reported to have had anal sex in the last 6 months, and were at least 18 years of age, from the sexual health and antiretroviral therapy centres. Structured questionnaires were administered, and blood samples were obtained to confirm HIV status. Anal swabs were collected for HPV-DNA detection and typing. MAIN OUTCOME MEASURES: The primary outcome was the prevalence of 'HPV-DNA infection'. The prevalence ratios (PR) were calculated using Cox proportional hazard model algorithms to analyse the association between exposure variables and HPV-infection. RESULTS: Complete data were available for 298 MSM and transgender women (HIV +n=131; HIV-n=167). The overall HPV-DNA prevalence was 65.1% and was higher in participants living with HIV as compared with HIV-negative (87% vs 48%; χ2p≤0.001). Likewise, 28.9% of participants living with HIV were infected with two or more than two types of HPV as compared with 18.8% participants without HIV(χ2 p≤0.001). The most frequent HPV type was HPV6/11 (46.9%), followed by HPV16 (35.1%), HPV18 (23.2%) and HPV35 (21.1%). HIV status (PR 2.81, 95% CI 2.16 to 3.82) and never condom use (PR 3.08, 95% CI 1.69 to 5.60)) were independently associated with prevalence of 'anal-HPV16 infection' when adjusting for confounding for age, other sexual and behavioural factors, for example, smoking and alcohol consumption. CONCLUSION: High prevalence of HPV indicates a substantial future risk of anal cancer in Pakistani MSM and transgender women, and particularly in those living with HIV. Current findings support anal Pap-smear HPV screening for this particular group and vaccination efforts for future generations.

Bioherbicidal ability and weed management of allelopathic methyl esters from Lantana camara.

Allelochemicals are secondary metabolites which are not edible and can be used as growth regulators and bio-herbicides. The goal of current study was to assess allelopathic ability of Lantana camara (Sage-plant) flowers against weeds viz. Avena fatua (Wild oat), Euphorbia helioscopia (Sun-spurge), Chenopodium album (Goosefoot), Phalaris minor (Canary-grass), and Rumex dentatus (Knotweed). Bioassay analysis of three methanolic fractions of the Combiflash from L. camara was performed at 50%, 75% and 100% concentration using germination percentage parameters, inhibition of plumule and radicle size. The fraction II of Combiflash strongly suppressed all weeds with negligible effect on T. aestivum. Gas chromatography-mass spectroscopy was conducted for the fraction, and isolated compounds were used to perform bioassays. From fraction II GC-MS detected four methyl esters of allelopathic fatty acid viz. Methyl oleate, methyl palmitate, methyl stearate and methyl linoleate. The evaluation of physiological effects of the bioassay revealed substantial suppression of chlorophyll, antioxidant enzymes (superoxide, dismutase peroxidase) and protein material in all weeds by methyl palmitate. Bioassay activity and study of physiological parameters revealed that the effective bio-herbicidal compound in Lantana camara flowers is methyl palmitate. This is the first time that methyl palmitate (a fatty acid methyl ester) has been related to herbicidal activity in L. camara flowers. It is proposed that field studies based on hormesis research and the mechanism of action of this compound be carried out.

In Silico Validation, Fabrication and Evaluation of Nano-Liposomes of Bistorta amplexicaulis Extract for Improved Anticancer Activity Against Hepatoma Cell Line (HepG2).

BACKGROUND: Bistorta amplexicaulis of the genus Polygonum (Polygonaceae) has been reported for its antioxidant and anticancer activities. However, the low cellular uptake of the compounds in its extract limits its therapeutic application. OBJECTIVES: The present study aimed at developing a nanoliposomal carrier system for B. amplexicaulis extracts for improved cellular uptake, thus resulting in enhanced anticancer activity. METHODS: Ultra Pressure Liquid Chromatography (UPLC) was used to identify major compounds in the plant extract. Nanoliposomes (NLs) were prepared by employing a thin-film rehydration method using DPPC, PEG2000DSPE and cholesterol, followed by characterization through several parameters. In vitro screening was performed against breast cancer cell line (MCF-7) and Hepatocellular carcinoma cell line (HepG-2) using MTT-assay. Raw extract and nanoliposomes were tested on Human Umbilical Vein Endothelial Cells (HUVEC). Moreover, molecular docking was performed to validate the data obtained through wet lab. RESULTS: The UHPLC method identified gallic acid, caffeic acid, chlorogenic acid and catechin as the major compounds in the extract. The NLs with a size ranging between 140-155 nm, zeta potential -16.9 to -19.8 mV and good polydispersity index of < 0.1 were prepared, with a high encapsulation efficiency of 81%. The MTT assay showed significantly (p > 0.05) high uptake and cytotoxicity of NLs as compared to the plant extract. Moreover, reduced toxicity against HUVEC cells was observed as compared to the extract. Also, the docking of identified compounds suggested a favorable interaction with the SH2 domain of both STAT3 and STAT5. CONCLUSION: Overall, the results suggest NLs as a potential platform that could be developed for the improved intracellular delivery of plant extract, thus increasing the therapeutic outcomes.

Assessing the Degree of Acute Esophageal Injury Secondary to Corrosive Intake: Insights From a Public Sector Hospitals of a Developing Country.

Background Caustic ingestion continues to be a significant problem worldwide especially in developing countries and particularly in the age group of under six years. Ingestion of caustic substances is a medical emergency in both the adult and pediatric population and is associated with high morbidity and mortality. The ingestion of caustic substances induces an extensive spectrum of injuries to the aerodigestive tract, which includes extensive necrosis and perforation of the esophagus and stomach. Objectives The main aims were to determine upper and lower esophageal injuries associated with corrosive intake and to compare esophageal injury with age and gender. Rationale Once we'll find the extent and severity of esophageal injury associated with corrosive intake within 24 hours, we'll be able to manage the case timely and to limit further complications and disabilities. Materials and Methods This descriptive cross-sectional study was conducted on 150 patients who presented with corrosive ingestion and underwent urgent endoscopic evaluation. Data were collected using self-designed pro forma. Endoscopic findings were classified according to the Zargar classification. A descriptive analysis of study variables was performed using SPSS v.21.0 (IBM Corp., Armonk, NY, USA). The chi-square test was used, and a p-value of less than 0.05 was considered statistically significant. Results Out of 150 patients under study, 103 (68.7%) were females and 47 (31.3%) were males. The most prevalent age group presenting with corrosive intake was found to be between 21 and 34 years of age (43.3%) in both genders. The most common part of the esophagus prone to corrosive insult is the upper esophagus (99.3%), whereas, regarding severity, the lower esophagus has more severe injuries (predominant being stage 2B, i.e., 32%). There are no statistically significant differences in esophageal injuries in different age groups (upper esophageal injury: 0.319; lower esophageal injury: 0.696) and genders (upper esophageal injury: 0.769; lower esophageal injury: 0.752).  Conclusions Most of the patients under study belong to the female gender and teen and younger age group. The predominant upper esophageal injury as a result of corrosive intake is stage 0 injury, and the least common is found to be stage 1 injury. The predominant lower esophageal injury as a result of corrosive intake is stage 2B injury, whereas the least common is found to be stage 4 injury.

Tuberculosis Resistance and Nanoparticles: Combating the Dual Role of Reactive Oxygen Species in Macrophages for Tuberculosis Management.

Increasing drift in antimicrobial therapy failure against Mycobacterium tuberculosis, the causative agent of tuberculosis (TB), and the advent of extended resistant strains strongly demand discovery of mechanisms underlying development of drug resistance. The emergence of resistance against anti-TB drugs has reached an alarming level in various parts of the world, providing an active platform for the design of new targeted drug delivery. Reactive oxygen species (ROS) have an important role in controlling TB pathogenesis. At macrophage activation, ROS that are produced inside macrophages directly kill resident bacteria. These ROS possess a dual character because they can kill macrophages along with the resident bacteria. Targeting these ROS can play a remarkable part in overcoming resistance of conventional drugs. Nanoparticles (NPs) have evolved as a potential drug carrier for targeted delivery and elimination of various resistance mechanisms against antimicrobials. Receptor-mediated targeting of macrophages via different NPs may be a promising strategy for combating drug resistance and enhancing efficacy of old-fashioned antimycobacterial agents.

DNA electromagnetic properties and interactions -An investigation on intrinsic bioelectromagnetism within DNA.

The question whether intrinsic bioelectromagnetism exists within DNA or not is an important and so far unexplored area of biology. We carried out a study of isolated genetic material, utilizing both prokaryotic and eukaryotic DNA, to measure any possible intrinsic electromagnetic effects or fields emanated within the molecules. Studies were carried out with extremely sensitive ultra-low-noise trans-impedance amplifiers and a high-precision data acquisition system to record any possible faintest electromagnetic signals from the concentrated, as well as diluted DNA, in vitro. Some experiments were performed to investigate any possible electromagnetic effects of high-frequency (HF) RF fields on the DNA under test. However, after extensive testing and careful measurements, we failed to detect any possible intrinsic or induced electromagnetic activity from the DNA as compared to simple water or empty chambers. We reached a conclusion that there does not seem to be any measurable intrinsic electromagnetic activity or fields present in the DNA material, whether in concentrated or diluted form, and if there were, any such activity or fields would be extremely minuscule to be detected with scientific precision by current human measurement methods.

Folate grafted thiolated chitosan enveloped nanoliposomes with enhanced oral bioavailability and anticancer activity of docetaxel.

Folate grafted and thiolated chitosan was synthesized and wrapped on the surface of mixed phosphatidylcholine based nanoliposomes (NLs) to improve the oral absorption and targeted pharmacological activity of anti-cancer drugs against breast cancer. In this study, a chitosan derived thiomer, having intrinsic properties of P-glycoprotein (P-gp) efflux pump inhibition, mucoadhesion and controlled drug release at a target site, was exploited to improve the performance of docetaxel (DTX) loaded NLs for better oral pharmacokinetics, targeted anti-cancer activity, liposomal stability and the physical characteristics of NLs. Thiomer enveloped nanoliposomes (ENLs) and bare nanoliposomes (NLs) were synthesized with the ingredient ratio pre-determined via Response Surface Methodology (RSM) plots by Design Expert® software. ENLs and NLs were thoroughly characterized for their surface chemistry, particle size, zeta potential, PDI, encapsulation efficiency, stability and release profile. ENLs were spherical in shape with a particle size of 328.5 ± 30 nm, a positive zeta potential of 18.81 ± 2.45 and a high encapsulation efficiency of 83% for DTX. Controlled release of DTX from formulations was observed for over 72 h for each formulation. The presence of thiol groups at the surface of the ENLs resulted in higher swelling and in situ gelling properties compared to the corresponding NLs. Furthermore, ENL/mucin mixtures showed a time dependent increase in viscosity for up to 12 h, leading to a 19.07-fold increased viscosity. Ex vivo permeation and P-glycoprotein inhibiting properties, studied in rat's small intestine, showed a 9.6-fold higher permeation and 13-fold enhancement of DTX in the presence of ENLs. In vitro cytotoxicity studies indicated that the ENLs can efficiently kill MD-MB-231 breast cancer cells with 200 fold lower IC50 values than DTX alone as a positive control. The pharmacokinetic study revealed that the ENLs significantly improved the oral bioavailability of DTX i.e. up to 13.6 fold as compared to an aqueous dispersion of DTX. Therefore, these enveloped hybrid nanoliposomes (ENLs) have the potential to be developed as useful nanocarriers for efficient oral delivery and breast cancer management using DTX.

Evaluation of antioxidant potential and HPLC based identification of phenolics in Polygonum amplexicaule extract and its fractions.

There is a growing interest for the plant-based medicines in pharmaceutical industry. Plant derived Antioxidants have gained huge importance regarding their medicinal value. The present study was designed to establish pharmaceutical value of Polygonum amplexicaule for their antioxidant activity using shoot, leaf and rhizome crude methanolic extract along with their n-butanolic, ethanolic, ethyl acetate and aqueous fractions. DPPH assay was used to assess antioxidants, which shows the maximum activity by crude methanolic extract of leaves (CMEL) having IC(50) 1.03 µg/ml where all other fractions showed IC(50) in a range of 1.03-58.2 µg/mL. The DNA plasmid protection assay showed that 10 ppm and 100 ppm concentrations of crude methanolic extracts (rhizome and leaf), aqueous fractions (shoot and leaf extract), n-butanolic fractions (shoot and leaf extract) and ethanolic fraction (rhizome extract) have DNA protection properties. TLC and HPLC based Identification of different antioxidants present in shoot, leaf and rhizome crude extracts and their fractions showed the presence of gallic acid, quercetin, catechin, caffeic acid, rutin, myricetin and kaempferol. This study suggested that this plant have high content of antioxidants, which needs to be investigated further for their medicinal and/cosmaceutical applications.

Hepatitis B virus among maintainence haemodialysis patients: a report from Karachi, Pakistan.

OBJECTIVE: To study the characterstics, pattern of viral markers, hepatic transaminases and other associated hepatitis virus infections among maintainence haemodialysis patients. METHOD: This prospective cross-sectional study was conducted at the dialysis unit of Sindh Institute of Urology and Transplantation Karachi on end-stage renal disease (ESRD) patients undergoing maintainence haemodialysis from November 2009 to April 2010. Patients on anti-viral drugs were excluded Blood samples were taken, immediately before start of dialysis, for Hepatitis B virus serology, anti-HCV, polymerase chain reaction (PCR) for HBV DNA and biochemical tests. Abdominal ultrasound was performed for liver, spleen, portal vein diameter and presence or absence of ascites. Data analysis was done by using SPSS 10.0. The level of significance was taken as 0.05. RESULTS: Out of 1220 ESRD patients on maintainence haemodialysis at SIUT, 124 were HBsAg positive but 26 patients were excluded as they had received anti-viral therapy. Finally 98 patients including 71 (72%) males and 27 (28%) females completed the study. The mean age was 34.98 +/- 12.67 years. Most of the patients did not have hepatitis symptoms. ALT level was raised above cutoff value of 20 IU/ml in 62.2% patients while AST was raised in 75.4% patients. HBeAg was positive in 34.6%, anti-HBe antibody was positive in 65% patients and HBV DNA was detected in 65.3%. More than half of the patients had HCV co-infection. Six patients had cirrhosis. Thirty four patients were non-replicating carriers. The mean duration of dialysis and duration of HBsAg positivity were significantly longer in those patients who had hepatitis B and hepatitis C coinfection (p<0.05). CONCLUSION: Hepatitis B virus infection in dialysis dependent patients is mostly asymptomatic. Mild transaminase elevation is frequently encountered.

Transcutaneous carbon dioxide pressure monitoring in a specialized weaning unit.

OBJECTIVE: To evaluate transcutaneously measured P(CO(2)) (P(tcCO(2))) values during ventilator weaning and during bronchoscopies on ventilated patients, and to compare P(tcCO(2)) values to P(aCO(2)) values from arterial blood analysis and end-tidal P(CO(2)) (P(ETCO(2))) values from capnography. METHODS: In our specialized weaning unit we measured P(tcCO(2)) in tracheostomized patients with prolonged weaning failure during daytime spontaneous breathing trials (SBTs) (23 measurement sessions in 15 patients), during their first nights off the ventilator (12 measurement sessions in 12 patients), during bronchoscopy while ventilated (80 measurement sessions in 21 patients), simultaneous with arterial blood draw for blood gas analysis (48 measurements in 38 patients), and simultaneous with P(ETCO(2)) measurements (39 measurements in 31 patients). RESULTS: There were often large changes (> 10 mm Hg) in P(tcCO(2)) during daytime SBTs (23%) and the initial overnight off-the-ventilator periods (42%), which influenced the decisions of whether to continue the SBT. P(tcCO(2)) often rose during bronchoscopy (mean +/- SD increase of 10.7 +/- 5.8 mm Hg), which influenced the physician to change the ventilator settings 44% of the time. P(aCO(2)) closely matched P(tcCO(2)) (mean +/- SD difference of 0.5 +/- 4.1 mm Hg). There was a greater difference between P(aCO(2)) and P(ETCO(2)) (3.7 +/- 7.7 mm Hg during prolonged exhalation, and 6.8 +/- 7.2 mm Hg during tidal breathing). CONCLUSIONS: Monitoring P(tcCO(2)) is very helpful in assessing and managing patients undergoing SBTs, during the first night off the ventilator, and during bronchoscopy on ventilated patients. P(tcCO(2)) more closely matches P(aCO(2)) than does P(ETCO(2)).