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Background: The harmonic scalpel, commonly used in laparoscopic surgery, now has promise for MRM dissection. The harmonic scalpel's high frequency mechanical vibrations cut and coagulate intraoperatively at the same time, generating less heat injury than electrocautery. Objective: To compare the outcome of modified radical mastectomy using harmonic scalpel versus electrocautery at tertiary care teaching hospital. Methodology: The randomized controlled trial was conducted from 12th September 2018 11th March 2019 at Department of Surgery, Liaquat University of Medical and Health Science, Jamshoro. A total of 128 patients with infiltrating ductal carcinoma undergoing modified radical mastectomy, 20 to 50 years of age were included. Chronic Hepatitis, diabetes, and neo-adjuvant treatment patients were excluded. Modified radical mastectomy employing harmonic scalpel for Group A and electrocautery for Group B. Postoperative problems such as seroma development, postoperative hematoma, marginal necrosis, lymphedema, and wound infection were also observed. Results: The mean age of women in group A was 39.81 ± 6.73 years and in group B was 39.45 ± 6.60 years. Mean duration of disease was 5.41 ± 1.91 months. The frequency of seroma formation in harmonic scalpel group as 7.81% vs 26.56% in electrocautery group, p=0.005), frequency of pain is 53.13% vs 68.75% respectively, p=0.070, frequency of hematoma is 1.56% vs 17.19% respectively, p=0.002), frequency of marginal necrosis was 0.0% vs 7.81% respectively, p=0.023), lymphedema 3.13% vs 14.06% respectively, p=0.027) and wound infection 17.19% vs 35.94% respectively, p=0.016. Conclusion: This research indicated that harmonic scalpel is superior to electrocautery in modified radical mastectomy.

Background: Herbal medicine is becoming increasingly popular among the general population. Before consulting a medical practitioner, patients already try home remedies. Therefore, it becomes more pertinent that the medical students as future doctors, must possess some knowledge of the herbal medicine. In Saudi Arabia, no study has yet evaluated this aspect in medical students. Methods: A cross-sectional study was performed with 240 medical students at a Health Sciences University. The survey questionnaire was validated, and descriptive/inferential statistics analyses were conducted by SPSS. Results: Data showed 50/240 students (20.8 %) used herbal medicines and majority 36 (72%) used it without consultation with doctors. Moreover, male understanding of the sources of herbal medicine was less compared to females' (p < 0.05). In terms of attitude, majority agreed with the benefits of herbal medicines in the maintenance and promotion of health (123, 51.3%) and to treat illness (127, 52.9%). However, medical students did not want to use herbal medicines (122, 50.8%) or recommend it to their families (161, 67.1%). A significant number of the respondents (160, 66.7%) opined that the herbal medicines could be taken with conventional or allopathic medications. Male students were significantly more inclined towards use of herbal medicine than their female counterparts (p < 0.05). Conclusion: Medical students were found to be unaware of some important aspects regarding herbal medicines, like use of herbs with drugs without consultation. Inclusion of proper courses on alternative medicine in medical curriculum can fill this gap and clarify the misconceptions among medical students.

The cell‑killing potential of most chemotherapeutic agents is enhanced by a temperature elevation. Isofraxidin (IF) is a coumarin compound widely found in plants, such as the Umbelliferae or Chloranthaceae families. IF induces anticancer effects in lung and colorectal cancer. To the best of our knowledge, the combined effects of hyperthermia (HT) and IF on heat‑induced apoptosis have not been reported. Acute monocytic leukemia U937 cells were exposed to HT with or without IF pre‑treatment. Apoptosis was measured by Annexin V‑FITC/PI double staining assay using flow cytometry and cell viability was observed by cell counting kit assay, DNA fragmentation. The mechanism involved in the combination was explored by measuring changes in the mitochondrial membrane potential, (MMP), intracellular ROS generation, expression of apoptosis related protein, and intracellular calcium ion level. It was demonstrated that IF enhanced HT‑induced apoptosis in U937 cells. The results demonstrated that combined treatment enhanced mitochondrial membrane potential loss and transient superoxide generation increased protein expression levels of caspase‑3, caspase‑8 and phosphorylated‑JNK and intracellular calcium levels. Moreover, the role of caspases and JNK was confirmed using a pan caspase inhibitor (zVAD‑FMK) and JNK inhibitor (SP600125) in U937 cells. Collectively, the data demonstrated that IF enhanced HT‑induced apoptosis via a reactive oxygen species mediated mitochondria/caspase‑dependent pathway in U937 cells.

News media always pursue informing the public at large. It is impossible to overestimate the significance of understanding the semantics of news coverage. Traditionally, a news text is assigned to a single category; however, a piece of news may contain information from more than one domain. A multi-label text classification model for news is proposed in this paper. The proposed model is an automated expert system designed to optimize CNN’s classification of multi-label news items. The performance of a CNN is highly dependent on its hyperparameters, and manually tweaking their values is a cumbersome and inefficient task. A high-level metaheuristic optimization algorithm, spotted hyena optimizer (SHO), has higher advanced exploration and exploitation capabilities. SHO generates a collection of solutions as a group of hyperparameters to be optimized, and the process is repeated until the desired optimal solution is achieved. SHO is integrated to automate the tuning of the hyperparameters of a CNN, including learning rate, momentum, number of epochs, batch size, dropout, number of nodes, and activation function. Four publicly available news datasets are used to evaluate the proposed model. The tuned hyperparameters and higher convergence rate of the proposed model result in higher performance for multi-label news classification compared to a baseline CNN and other optimizations of CNNs. The resulting accuracies are 93.6%, 90.8%, 68.7%, and 95.4% for RCV1-v2, Reuters-21578, Slashdot, and NELA-GT-2019, respectively.

Due to the exponential increase in internet and social media users, fake news travels rapidly, and no one is immune to its adverse effects. Various machine learning approaches have evaluated text and images to categorize false news over time, but they lack a comprehensive representation of relevant features. This paper presents an automated method for detecting fake news to counteract the spread of disinformation. The proposed multimodal EFND integrates contextual, social context, and visual data from news articles and social media to build a multimodal feature vector with a high level of information density. Using a multimodal factorized bilinear pooling, the gathered features are fused to improve their correlation and offer a more accurate shared representation. Finally, a Multilayer Perceptron is implemented over the shared representation for the classification of fake news. EFND is evaluated using a group of standard fake news datasets known as “FakeNewsNet”. EFND has outperformed the baseline and state-of-the-art machine learning and deep learning models. Furthermore, the results of ablation studies have demonstrated the efficacy of the proposed framework. For the PolitiFact and GossipCop datasets, the EFND has achieved an accuracy of 0.988% and 0.990%, respectively.

Five medicinal plants Mentha piperita L., Trachyspermum ammi L., Viola odorata Linn., Matricaria chamomilla L. and Foeniculum vulgare Mill. were selected for their in vitro and in vivo evaluation of anti-Helicobacter pylori activity. In vitro evaluation was performed by using disk diffusion method and minimum inhibitory concentrations were noted while rat models were selected for in vivo activity against four Helicobacter pylori strains isolated form gastric mucosa. Mentha piperita showed largest zone of inhibition with 9 mm diameter among all other extracts. All the plants showed promising anti-Helicobacter pylori activity against four isolates and a reference strain at concentrations of 125, 250, 500 and 1000 μg/ml in comparison with Amoxicillin 1 μg/ml but least MIC was exhibited by Mentha piperita followed by in vivo testing where it competed Amoxicillin at 1000 mg/kg by achieving 80% eradication of Helicobacter pylori in mucosa of infected rats justified by histological examination of stomach. It was concluded that medicinal plants possess strong anti-Helicobacter pylori activity and can be considered a potential source of safe and effective alternative regimens for the eradication of Helicobacter pylori.

BackgroundIn the UK, the National Health Service has various incentivisation schemes in place to improve the provision of high-quality care. The Quality Outcomes Framework (QOF) and other Pay for Performance (P4P) schemes are incentive frameworks that focus on meeting predetermined clinical outcomes. However, the ability of these schemes to meet their aims is debated.Objectives(1) To explore current incentive schemes available in general practice in the UK, their impact and effectiveness in improving quality of care and (2) To identify other types of incentives discussed in the literature.MethodsThis systematic literature review was conducted using the Preferred Reporting Items for Systematic Reviews and Meta-Analyses guidelines. Six databases were searched: Cochrane, PubMed, National Institute for Health and Care Excellence Evidence, Health Management Information Consortium, Embase and Health Management. Articles were screened according to the selection criteria, evaluated against critical appraisal checklists and categorised into themes.Results35 articles were included from an initial search result of 22087. Articles were categorised into the following three overarching themes: financial incentives, non-financial incentives and competition.DiscussionThe majority of the literature focused on QOF. Its positive effects included reduced mortality rates, better data recording and improved sociodemographic inequalities. However, limitations involved decreased quality of care in non-incentivised activities, poor patient experiences due to tick-box exercises and increased pressure to meet non-specific targets. Findings surrounding competition were mixed, with limited evidence found on the use of non-financial incentives in primary care.ConclusionCurrent research looks extensively into financial incentives, however, we propose more research into the effects of intrinsic motivation alongside existing P4P schemes to enhance motivation and improve quality of care.

Cadmium (Cd) is a heavy metal and a highly toxic pollutant that is released into the environment as a byproduct of most modern factories and industries. Cd enters our body in significant quantities from contaminated water, cigarette smoke, or food product to many detrimental health hazards. Based on causal association all the Cd-related or derived compounds have been classified as carcinogens. In this study, we present an overview of the published literature to understand the molecular mechanisms for Cd-induced carcinogenesis and its prevention. In acute Cd poisoning production of reactive oxygen species is a key factor. However, chronic Cd exposure can transform cells to become more resistant to oxidative stress. Also, as an epigenetic mechanism Cd acts indirectly on DNA repair mechanisms via alteration of reactions upstream. Those transformed cells acquire resistance to apoptosis and deregulation of calcium homeostasis. Leading to uncontrolled carcinogenic cell proliferation and inherent DNA lesions. Flavonoids commonly found in plant foods have been shown to have a protective effect against Cd-induced carcinogenicity. A wide variety of tumorigenic mechanisms involved in chronic Cd exposure and the beneficial effects of flavonoids against Cd-induced carcinogenicity necessitate further investigations.

Colorectal cancer (CRC) is one of the leading causes of mortality among men and women. Chemo-resistance, adverse effects and disease recurrence are major challenges in the development of effective cancer therapeutics. Substantial literature on this subject highlights that populations consuming diets rich in fibers, fruits and vegetables have a significantly reduced incidence rate of CRC. This chemo-preventive effect is primarily associated with the presence of phytochemicals in the dietary components. Plant-derived chemical agents act as a prominent source of novel compounds for drug discovery. Phytochemicals have been the focus of an increasing number of studies due to their ability to modulate carcinogenic processes through the alteration of multiple cancer cell survival pathways. Despite promising results from experimental studies, only a limited number of phytochemicals have entered into clinical trials. The purpose of the current review is to compile previously published pre-clinical and clinical evidence of phytochemicals in cases of CRC. A PubMed, Google Scholar and Science Direct search was performed for relevant articles published between 2008-2018 using the following key terms: 'Phytochemicals with colorectal cancers', 'apoptosis', 'cell cycle', 'reactive oxygen species' and 'clinical anticancer activities'. The present review may aid in identifying the most investigated phytochemicals in CRC cells, and due to the limited number of studies that make it from the laboratory bench to clinical trial stage, may provide a novel foundation for future research.

Introduction: Various drug–food interactions exist that may hinder treatment and can sometimes be lethal. Our aim was to assess the level of public knowledge and awareness in Jeddah city, Western Saudi Arabia, about drug–food interactions, along with the effects of demographics on their knowledge. Methods: A survey questionnaire was administered in this cross-sectional study to participants spread across multiple locations in Jeddah, including in malls and public gatherings. Participants included both males and females. Sample size was calculated through Raosoft® software. Data analysis was executed using IBM Statistic SPSS and the level of statistical significance was set at p &lt, 0.05. Results: A total of 410 people participated in the study and only 92.68% (380) of responses were enrolled in the study, 7.32% (30) were not enrolled due to the exclusion criteria. Surprisingly, only six out of eighteen questions regarding drug–food interactions in the administered questionnaire were correctly answered by 380 participants. Data indicated that the participants had a poor to intermediate level of both knowledge and awareness with respect to drug–food interactions. Furthermore, participants showed moderate to strong awareness of the effects of alcohol and tea generally, and their interaction with medication. Conclusion: Participants in our study showed inadequate knowledge of basic and fundamental information about drug–food interactions, which highlights the dire need to increase awareness.

Protein homeostasis or proteostasis, the correct balance between production and degradation of proteins, is an essential pillar for proper cellular function. Among the several cellular mechanisms that disrupt homeostatic conditions in cancer cells, hyperthermia (HT) has shown promising anti-tumor effects. However, cancer cells are also capable of thermoresistance. Indeed, HT-induced protein denaturation and aggregation results in the up regulation of heat shock proteins, a group of molecular chaperones with cytoprotective and anti-apoptotic properties via stress-inducible transcription factor, heat shock factor 1(HSF1). Heat shock proteins assist in the refolding of misfolded proteins and aids in their elimination if they become irreversibly damaged by various stressors. Furthermore, HSF1 also initiates the unfolded protein response in the endoplasmic reticulum (ER) to assist in the protein folding capacity of ER and also promotes the translation of pro-survival proteins' mRNA such as activating transcription factor 4 (ATF 4). Moreover, HT associated induction of microRNAs is also involved in thermal resistance of cancer cells via up-regulation of anti-apoptotic Bcl-2 proteins and down regulation of pro-apoptotic Bax and caspase 3 activities. Another cellular protection in response to stressors is Autophagy, which is regulated by the Mammalian target of rapamycin (mTOR) protein. Kinase activity in mTOR phosphorylates HSF1 and promotes its nuclear translocation for heat shock protein synthesis. Over-expression of heat shock proteins are reported to up-regulate Beclin-1, an autophagy initiator. Moreover, HT-induced reactive oxygen species (ROS) generation is sensitized by transcription factor NF-E2 related factor 2 (Nrf2) and activates the cellular expression of antioxidants and autophagy gene. Furthermore, ROS also potentiates autophagy via activation of Beclin-1. Inhibition of thermotolerance can potentiate HT-induced apoptosis. Here, we outlined that heat stress alters cellular proteins which activates cellular homeostatic processes to promote cell survival and make cancer cells thermotolerant.

BackgroundThe introduction of financial incentives, such as the quality and outcomes framework (QOF), historically lead to improvements in standardising practice. However, with shifting demands on healthcare providers, are these schemes still enough to drive high-quality care?AimTo explore current incentives, intrinsic and extrinsic, their role and effectiveness in improving quality of care and how they are perceived by GPs.MethodMixed methods study using two systematic literature reviews including 44 papers and 18 semi-structured interviews with GPs.ResultsIn the literature, QOF was associated with reduced socioeconomic inequalities, decreased mortality and improved outcomes. However, the absence of control groups and the simultaneous analysis of multiple indicators complicates the findings. GPs agreed with the literature and viewed financial incentives as beneficial, however, they felt the key driver in providing good-quality care was their intrinsic motivation. Financial incentives were found to contribute to depersonalised care, diluted provision of non-incentivised activities and hindered overall practice. The results from the second literature review were in keeping with the views of the participants. They illustrated the importance of managing factors contributing to physician burnout, reduced performance, and low job satisfaction, which can result in the provision of low-quality care.ConclusionFinancial incentives have the potential to induce behaviour change, however, their use in quality improvement is limited when used alone. If used in an environment that nurtures intrinsic motivation, healthcare providers will be more driven to achieve a higher quality of care and will be better able to cope with shifting demands.

Heat has been used as a medicinal and healing modality throughout human history. The combination of hyperthermia (HT) with radiation and anticancer agents has been used clinically and has shown positive results to a certain extent. However, the clinical results of HT treatment alone have been only partially satisfactory. Cell death following HT treatment is a function of both temperature and treatment duration. HT induces cancer cell death through apoptosis; the degree of apoptosis and the apoptotic pathway vary in different cancer cell types. HT-induced reactive oxygen species production are responsible for apoptosis in various cell types. However, the underlying mechanism of signal transduction and the genes related to this process still need to be elucidated. In this review, we summarize the molecular mechanism of apoptosis induced by HT, enhancement of heat-induced apoptosis, and the genetic network involved in HT-induced apoptosis.

Very few empirical studies in migration and citizenship focus on immigrants' perceptions of the concept of citizenship and its prominent features. Based on the semi-structured interviews of 8 Pakistani immigrants (Swedish citizenship holders and non-citizenship holders) in Sweden, this study aimed to understand how these immigrants perceive and define Swedish citizenship, and how do they value it in material, symbolic, and emotional terms. Through conceptual framework and literature review of citizenship in its symbolic, materialistic, and emotional term, three central approaches mobility, sense of security, and sense of belonging were derived and analyzed. The findings of this research suggest that immigrants who participated in this study tend to value Swedish citizenship. They are very concerned about their security and protection which is provided by the state to a citizen in every sphere of life; economically, socially, politically; protection against the deportation; and while traveling anywhere in the world. These immigrants perceived that passport makes a difference, and during travel, it enables and constrains inequalities, restrictions, and uncertainties in the mobility arena. Furthermore, when it comes to belongings: the interviewee relate it to work, spending time in the country, and participation in the society. Finally, this study recommends further research with different migrants groups in Sweden to understand the true essence of Swedish citizenship in immigrants' worldview.

Helicobacter pylori is involved in the pathogenesis of gastritis, peptic ulcer, and gastric cancer. The infection is prevalent in more than half of the world's population. Although the infection may lead to detrimental consequences, still the majority of the infected individuals only develop mild gastritis. Several factors are behind this paradoxical outcome including virulence of the infecting H. pylori strains, genetic background of the host, and factors related to lifestyle such as dietary habits. Among these, lifestyle including dietary factors was not in the limelight, until recently, as one of the important factors that could modulate H. pylori-linked gastric diseases. This review is directed to gather and elucidate the role of dietary components in augmenting or attenuating pathological processes initiated by H. pylori. Available evidence strongly supports the notion that the diet may play a critical role in defining the final outcome of H. pylori infection particularly if certain dietary components are taken on a regular basis for a long time. Despite a recent surge in research related to the role of dietary ingredients, further studies involving large-scale clinical trials are required to gain a better understanding of the precise role played by the dietary ingredients in H. pylori-associated pathogenesis.

Shikonin (SHK), a natural naphthoquinone derived from the Chinese medical herb Lithospermum erythrorhizon, induces both apoptosis and necroptosis in several cancer cell lines. However, the detailed molecular mechanisms involved in the initiation of cell death are still unclear. In the present study, caspase-dependent apoptosis was induced by SHK treatment at 1μM after 6h in U937 cells, with increase in DNA fragmentation, generation of intracellular reactive oxygen species (ROS), fraction of cells with low mitochondrial membrane potential (MMP), and in the expression of BH3 only proteins Noxa and tBid. Interestingly, caspase-independent cell death was also detected with SHK treatment at 10μM, observed as increase in SYTOX® Green staining and release of lactate dehydrogenase (LDH). Necrostatin-1 (Nec-1) completely inhibited the SHK-induced leakage of LDH and SYTOX® Green staining. Cell permeable exogenous glutathione (GSH) completely inhibited 1μM SHK-induced apoptosis and converted 10μM SHK-induced necroptosis to apoptosis. Gene expression profiling revealed that 353 genes were found to be significantly regulated by 1μM and 85 genes by 10μM of SHK treatment, respectively. Among these genes, the transcription factor 3 (ATF3) and DNA-damage-inducible transcript 3 (DDIT3) were highly expressed at 1μM of SHK treatment, while tumor necrosis factor (TNF) expression mainly increased at 10μM treatment. These findings provide novel information for the molecular mechanism of SHK-induced apoptosis and necroptosis.

Transforming growth factor-β-activated kinase1 (TAK1) plays an anti-apoptotic role in response to multiple stresses. TAK1 inhibitor, 5Z-7-oxozeaenol (OZ) has been studied for its apoptotic effects. However, the combined effect of OZ with physical stresses remains to be elusive. Therefore, in this study we focussed to determine the combined effects of OZ with hyperthermia (HT) using Molt-4 cell line.Molt-4 cells were pre-treated with OZ for 1 h followed by heat exposure (44 °C, 10 min) and harvested 24 h after incubation at 37 °C, apoptosis was measured by Annexin V-FITC/PI double staining assay using flow cytometry and cell growth was observed by cell counting assay. Further mechanism involved in the combination was investigated by measuring mitochondrial membrane potential (MMP), intracellular ROS generation, expression of apoptosis related protein, intracellular calcium ion level and Fas activity.Combination of OZ with HT significantly enhances MMP loss and superoxide generation. Furthermore, OZ pre-treatment promotes caspase-8 cleavage, Fas externalisation, caspase 3 activity and intracellular calcium ion levels. OZ pre-treatment decreased the expression of HT-induced Bcl-2 and increased the expression of pro-apoptotic Bax, while markedly suppressed the phosphorylation of JNK and p38. In addition, increased expression of CHOP following combined treatment indicates that ER stress may also involve in the enhancement of HT-induced apoptosis.Our data showed for the first time that OZ sensitizes Molt-4 cells to HT-induced apoptosis via extrinsic and intrinsic apoptotic pathways. Furthermore, ROS and ER stress may also play role in the enhancement of HT-induced apoptosis by OZ.

Cinobufotalin (CB), one of the bufadienolides prepared from toad venom, was investigated for its cytotoxicity, and the underneath mechanism involved. We primarily utilized DNA fragmentation assay and microscopic observation to assess the effect of various doses of CB in human lymphoma U937 cells. Following that, we investigated other parameters involved in cell death mechanism such as reactive oxygen species (ROS), mitochondrial membrane potential (MMP), and apoptotic proteins activation. HeLa cells were concomitantly used to generalize the data observed. Our results show that CB caused significant DNA fragmentation, decrease of MMP, and an increase in the intracellular Ca(2+) ion and ROS production. In addition, CB induced upregulation of Fas protein, proteolytic activation of cytochrome c, caspase-2, -3, -8 and -9 together with the activation of Bid and Bax. Our findings were further validated using either Fas/FasL antagonist or pan-caspase inhibitor to significantly inhibit CB-induced DNA fragmentation. In our study, we suggest that CB induces caspase dependent cell death in U937 cells, and that Fas plays a role in CB-induced apoptosis. Altogether, our data provides novel insights of the mechanism of action of CB and its potential as a future chemotherapeutic agent.

The catechins, a family of polyphenols found in tea, can evoke various responses, including apoptosis. In this study we investigated whether the chemical modification of (-)-epigallocatechin gallate (EGCG) could enhance its apoptosis activity. We found that one of the catechin conjugated with capric acid [(2R,3S)-3',4',5,7-tetrahydroxyflavan-3-yl decanoate; catechin-C10] was most potent to induce apoptosis in U937 cells. C10 treatment resulted in a significant increase in reactive oxygen species (ROS) formation, mitochondrial membrane potential (MMP) loss, cytochrome c release caspase-9 and caspase-3 activation. In addition to this C10 also activated extrinsic pathway significantly as evident by time-dependent increase in Fas expression and caspase-8 activity. C10 mediated cleavage of Bid may be an important event for cross talk between intrinsic and extrinsic signaling. Moreover, pre-treatment of cells with anti-oxidant N-acetyl-L-cysteine (NAC) significantly prevented C10-induced apoptosis but did not protect MMP loss. Treatment of cells with pan-caspase inhibitor significantly inhibited apoptosis indicating that caspases are playing key role. In addition to this C10 was found to induce apoptosis in human colon cancer (HCT116) cells while it showed resistance to human keratinocytes (HaCat). In short our results showed that the optimal fatty acid side chain length is required for the apoptosis inducing activity of catechin derivatives in U937 cells.

MDM2 is a critical negative regulator of the p53 tumor suppressor protein. Recently, nutlins, small-molecule antagonists of MDM2, have been developed to inhibit the p53-MDM2 interaction and activate p53 signaling. The expressions of DR4 and DR5, Tumor necrosis factor (TNF)-related apoptosis-inducing ligand (TRAIL) receptors, are regulated by p53. In this study, the combined effects of nutlin-3 and TRAIL on apoptosis were investigated in HOS and HCT116 cells, which express wild-type p53. Nutlin-3 and TRAIL synergistically enhanced apoptosis owing to their intrinsic and extrinsic pathway signals, respectively. The increase in the Bid expression level and the decrease in the expression levels of anti-apoptotic proteins, c-FLIP and XIAP, were involved in this apoptosis enhancement. Furthermore, nutlin-3 activated the DR5 promoter and increased the expression levels of DR5 at mRNA and protein levels. These results indicate that the combination, treated with nutlin-3 and TRAIL, is useful for apoptosis induction in malignant cells expressing wild-type p53.

Despite improvements in chemotherapy and surgery in the treatment of osteosarcoma (OS), satisfactory results are still difficult to achieve. Novel therapeutic modalities need to be developed for osteosarcoma treatment. The combined effects of tumor necrosis factor-related apoptosis-inducing ligand (TRAIL) and ionizing radiation (IR) on human OS cells were investigated. IR and TRAIL treatment synergistically decreased the cell viability and enhanced apoptosis in OS cell lines. IR pretreatment enhances TRAIL-induced Bid and caspase-3 activations. Decreases in the expression levels of the antiapoptotic proteins c-FLIP and XIAP also associated with apoptosis enhancement. Furthermore, IR pretreatment enhanced DR4 and DR5 expressions at the transcription stage. These results can become the basic lines of evidence for the future treatment of OS using TRAIL with IR.

The change in transfection efficiency of electroporation by the combined treatment with mild preheating (40 degrees C for 30 min) was investigated. HCT 116, HeLa S3 and SGC 7901 cells were treated with electroporation in medium containing pBKCMV-Luc plasmid with or without preheating. After 24 h, luciferase activity was increased by 36, 28 and 77%; luciferase mRNA transcription was increased by 45, 50 and 68%; and fluorescein isothiocyanate-dextran accumulation was increased by 9, 35 and 15% in preheated groups, respectively. These results demonstrate that the transfection efficiency was enhanced by mild preheating. The mechanism partially involves increased macromolecular particle accumulation.

The apoptosis-inducing ability of hybrid compounds composed of macrosphelide and thiazole-containing side chain of epothilones was investigated. Among the tested series of hybrid compounds the one containing thiazole side chain at C15 (MSt-2) showed the maximum potency to induce apoptosis, while another containing thiazole side chain at C3 (MSt-6) was less potent. MSt-2 was found to induce apoptosis in human lymphoma (U937) cells in a dose- and time-dependent manner as confirmed by DNA fragmentation analysis. MSt-2 treated cells showed rapid reactive oxygen species (ROS) formation and c-Jun N-terminal kinase (JNK) activation. Furthermore, significant activation of extrinsic pathway as evident by Fas expression and caspase-8 activation was also observed. MSt-2-mediated decreased expression of Bid is an important event for cross talk between intrinsic and extrinsic signaling. N-acetyl-l-cysteine pre-treatment rescued cells from MSt-2-induced ROS formation, mitochondrial membrane potential (Delta psi(m)) loss, Fas expression, caspase-8 and -3 activation and DNA fragmentation. Moreover, antioxidant enzymes catalase and/or superoxide dismutase conjugated with polyethylene glycol also inhibit MSt-2-induced ROS formation, apoptosis and Delta psi(m) loss suggesting thereby pro-oxidant effects of MSt-2. Furthermore, JNK and pan-caspase inhibitors also protect cells from MSt-2-induced apoptosis. In addition to this, MSt-2 was found to be more potent in human colon carcinoma (HCT116) and human gastric cancer (AGS) cells while it has no effect on human normal dermal fibroblast. The important structure-activity relationship observed in the current study which makes MSt-2 more potent than MSt-6 is the position of thiazole side chain and stereochemistry of position 3 in chemical structure. In short the results of our study demonstrate that MSt-2-induced rapid ROS generation and mitochondrial dysfunction in cells trigger events responsible for mitochondria-dependent apoptosis pathway.

Hyperthermia (HT) is considered to be a possible treatment modality for various cancers, and its pleiotropic effects support its combined use with radiotherapy and/or chemotherapy. However, clinical results by HT alone have not always been satisfactory. In mammalian cells, HT elicits a wide spectrum of alterations in cellular morphology, biochemistry and function. One of these HT-induced alterations, oxidative stress, has been attributed to the increased production of reactive oxygen spaces (ROS), and is known to play an important role as an intracellular mediator of HT-induced cell death, including apoptosis. Indeed, it has been well established that increases in intracellular oxidative stress significantly enhance HT-induced apoptosis. Attention has therefore been focused on the development of heat sensitizers to modulate the intracellular ROS. Interestingly, the modification of oxidative stress via addition of ROS-generating compounds significantly enhanced the apoptosis elicited by HT. In this chapter, we describe the induction of oxidative stress by HT and enhancement of HT-induced apoptosis by oxidative stress modification, and we discuss the possible mechanisms underlying this enhancement.

The combined effects of hyperthermia (44 degrees C, 20 min) or X-rays (10 Gy) and a new class of furan-fused tetracyclic synthesized compounds (DFs), on apoptosis in human lymphoma U937 cells were investigated. Among the tested compounds (DF1 approximately 6), the combined treatment of 10 microM DF with TIPS (triisopropylsilyloxy) (Designated #3 DF3) and hyperthermia showed the largest potency to induce DNA fragmentation at 6 h after hyperthermia but no enhancement was observed if it was combined with X-rays. Enhancement of hyperthermia-induced apoptosis by DF3 in a dose-dependent manner was observed. When the cells were treated first with DF3 at a nontoxic concentration of 20 microM, and exposed to hyperthermia afterwards, a significant enhancement of heat-induced apoptosis was evidenced by DNA fragmentation, morphological changes and phosphatidylserine externalization. The activation of Bid, but no change of Bax and Bcl-2 were observed after the combined treatment. The release of cytochrome c from mitochondria to cytosol, which was induced by hyperthermia, was enhanced by DF3. Mitochondrial transmembrane potential was decreased and the activation of caspase-3 and caspase-8 was enhanced in the cells treated with the combination. Externalization of Fas was observed following the combined treatment. Flow cytometry revealed rapid and sustained increase of intracellular superoxide due to DF3, and showed subsequent and transient increase in the formation of intracellular hydrogen peroxide (H(2)O(2)), which was further increased when hyperthermia was combined. These results indicate that the intracellular superoxide and H(2)O(2) generated by DF3 enhance the hyperthermia-induced apoptosis via the Fas-mediated mitochondrial caspase-dependent pathway.

Hyperthermia is a useful adjunct in cancer therapy as it can increase the effectiveness and decrease the toxicity of currently available cancer treatments such as chemotherapy and radiation. In the present study, we investigated whether 41 degrees C hyperthermia (mild HT) for 20 min can enhance macrosphelide (MS5)-induced apoptosis in human lymphoma U937 cells. Our results revealed that, compared with MS5 (5 microM) and mild HT alone, the combined treatment exhibited significant enhancement in apoptosis at 6 h, which was evaluated by observing morphological changes and DNA fragmentation. Marked increase in the reactive oxygen species (ROS) generation was observed immediately after the combined treatment. Significant increase in Fas externalization, caspase-8 and caspase-3 activation, and loss of mitochondrial membrane potential (MMP) was found after the combined treatment compared with MS5 and mild HT alone. Moreover, this combination can also alter the expression of apoptosis-related proteins as evident by the cleavage of Bid and down-regulation of Bcl-2 while no change in the expression of Bax was observed. Furthermore, an immediate rise in the intracellular calcium ion ([Ca(2+)]i) concentration was observed after the combined treatment, which continuously increased in a time-dependent manner. In addition, mild HT treatment alone also increases [Ca(2+)]i concentration without inducing apoptosis. Our data indicate that early increase in ROS generation is mainly responsible for the enhancement of apoptosis after the combined treatment.

Hybrid compounds of macrosphelides and epothilones, both of which are natural macrolides having a 16-membered skeleton, were designed and synthesized using a ring-closing metathesis (RCM) strategy. Some of these hybrids were found to exhibit notable apoptosis-inducing activity against human lymphoma cells with higher potency than parent natural macrosphelides, and to be a promising lead compound for development of a new antitumor agent.

The fundamental idea and the effects of heat on cancer cells are well known. However, the results obtained in therapy by hyperthermia (HT) alone have been only partially satisfactory. Treatment at temperatures between .40 and 44 degrees C is cytotoxic for cells in an environment with a low oxygen partial pressure and low pH, conditions that are found specifically within tumour tissue, due to insufficient blood perfusion. Under such conditions radiotherapy is less effective, and systemically applied cytotoxic agents will reach such areas in lower concentrations than in well-perfused areas. Therefore, clinically, it is preferred to use hyperthermia in combination with radiation therapy and chemotherapy. Hyperthermia can be applied by several methods: local hyperthermia by external or internal energy sources; regional hyperthermia by perfusion of organs or limbs, or by irrigation of body cavities; and whole-body hyperthermia. Number of studies have reported the combination of thermo-radiotherapy. Consequently, much attention has been focussed on identifying agents among the conventional chemotherapeutic substances that can sensitise tumour cells to hyperthermia-induced damage with minimal effects on normal cells. In this review, we overviewed important mechanisms of hyperthermia-induced apoptosis and the substances which can act as heat sensitisers in cancer therapy.

Environmental stress induces damage that activates an adaptive response in any organism. The cellular stress response is based on the induction of cytoprotective proteins, the so-called stress or heat shock proteins (HSPs). HSPs are known to function as molecular chaperones which are involved in the therapeutic approach of many diseases. Therefore in the current study we searched nontoxic chaperone inducers in chemical compounds isolated from medicinal plants. Screening of 80 compounds for their Hsp70-inducing activity in human lymphoma U937 cells was performed by western blotting. Five compounds showed significant Hsp70 up-regulation among them shikonin was most potent. Shikonin was able to induce Hsp70 at 0.1 µM after 3 h without activation of heat shock transcription factor 1 (HSF-1). It also induces significant reactive oxygen species generation. The expression level of genes responsive to shikonin was studied using global-scale microarrays and computational gene expression analysis tools. Significant increase in the nuclear factor erythroid 2-related factor 2 (Nrf2, NFEL2L2) -mediated oxidative stress response was observed that leads to the activation of HSP. The results of gene chip analysis were further confirmed by real-time qPCR assay. In short, the detailed mechanisms of Hsp70 induction by shikonin is not fully understood, Nrf2 and its target genes might be involved in the Hsp70 up-regulation in U937 cells.

Synthesis of macrosphelides containing a pyridyl or benzimidazolyl side chain is described. The synthesis was accomplished through sequential assembling of suitable chiral blocks and ring-closing metathesis (RCM) with a high efficiency. These novel derivatives were evaluated on the apoptosis-inducing activity against human lymphoma cells.

Despite improvements in chemotherapy and surgery in the treatment of osteosarcoma (OS), satisfactory results are still difficult to achieve. Novel therapeutic modalities need to be developed for osteosarcoma treatment. The combined effects of tumor necrosis factor-related apoptosis-inducing ligand (TRAIL) and ionizing radiation (IR) on human OS cells were investigated. IR and TRAIL treatment synergistically decreased the cell viability and enhanced apoptosis in OS cell lines. IR pretreatment enhances TRAIL-induced Bid and caspase-3 activations. Decreases in the expression levels of the antiapoptotic proteins c-FLIP and XIAP also associated with apoptosis enhancement. Furthermore, IR pretreatment enhanced DR4 and DR5 expressions at the transcription stage. These results can become the basic lines of evidence for the future treatment of OS using TRAIL with IR.

BACKGROUND Anomalous expression of activation-induced cytidine deaminase (AID) in Helicobacter pylori-infected gastric epithelial cells has been postulated as one of the key mechanisms in the development of gastric cancer. AID is overexpressed in the cells through nuclear factor (NF)-kappaB activation by H. pylori and hence, inhibition of NF-kappaB pathway can downregulate the expression of AID. Curcumin, a spice-derived polyphenol, is known for its anti-inflammatory activity via NF-kappaB inhibition. Therefore, it was hypothesized that curcumin might suppress AID overexpression via NF-kappaB inhibitory activity in H. pylori-infected gastric epithelial cells. MATERIALS AND METHODS MKN-28 or MKN-45 cells and H. pylori strain 193C isolated from gastric cancer patient were used for co-culture experiments. Cells were pretreated with or without nonbactericidal concentrations of curcumin. Apoptosis was determined by DNA fragmentation assay. Enzyme-linked immunosorbent assay was performed to evaluate the anti-adhesion activity of curcumin. Real-time polymerase chain reaction was employed to evaluate the expression of AID mRNA. Immunoblot assay was performed for the analysis of AID, NF-kappaB, inhibitors of NF-kappaB (IkappaB), and IkappaB kinase (IKK) complex regulation with or without curcumin. RESULTS The adhesion of H. pylori to gastric epithelial cells was not inhibited by curcumin pretreatment at nonbactericidal concentrations (< or =10 micromol/L). Pretreatment with nonbactericidal concentration of curcumin downregulated the expression of AID induced by H. pylori. Similarly, NF-kappaB activation inhibitor (SN-50) and proteasome inhibitor (MG-132) also downregulated the mRNA expression of AID. Moreover, curcumin (< or =10 micromol/L) has suppressed H. pylori-induced NF-kappaB activation via inhibition of IKK activation and IkappaB degradation. CONCLUSION Nonbactericidal concentrations of curcumin downregulated H. pylori-induced AID expression in gastric epithelial cells, probably via the inhibition of NF-kappaB pathway. Hence, curcumin can be considered as a potential chemopreventive candidate against H. pylori-related gastric carcinogenesis.

Inflammatory cytokine interleukin-8 (IL-8) and reactive oxygen species (ROS) overexpressed in the gastric mucosa when exposed to Helicobacter pylori, defined as a class I carcinogen. Moreover, infection with H. pylori leads to morphological changes in co-cultured cells known as hummingbird phenomenon along with increased motility. Resveratrol, a highly abundant polyphenol in red grapes, has shown anti-inflammatory, anti-cancer, cardioprotective and neuroprotective activities. However, the effect of resveratrol in H. pylori-infected cells has not been investigated. The present study was, therefore, aimed to evaluate the effect of resveratrol on the induction of IL-8, ROS and hummingbird morphology in H. pylori-infected gastric epithelial cells. The non-toxic concentration of resveratrol for both H. pylori and epithelial cells was determined by brucella broth dilution method and DNA fragmentation assay. The non-toxic resveratrol (< or =100 microM) treatment did not demonstrate any inhibitory effect against H. pylori adhesion to gastric epithelial cells. However, preincubation of the cells with 75 and 100 muM of resveratrol significantly (p