The Fluorescent Cell Line SW620-GFP Is a Valuable Model to Monitor Magnetic Hyperthermia.

In this work, the cell line SW620-GFP has been used in a complete magnetic hyperthermia assay, from the preparation of the ferrofluid with folate-coated iron oxide nanoparticles to in vivo experiments. The physical and chemical characterization of the nanoparticles evidenced their superparamagnetic behaviour, an average diameter of 12 ± 4 nm, a 2 nm coat thickness, and a high-power loss density. The main innovation of the work is the exclusive capability of viable SW620-GFP cells to emit fluorescence, enabling fast analysis of both, cell viability in vitro with an epifluorescence microscope and tumour size and shape in vivo in a non-invasive manner using the iBox technology. Moreover, with this imaging technique, it was possible to demonstrate the successful tumour size reduction in mice applying magnetic hyperthermia three times a week over 3 weeks.

Dry but Not Humid Thermal Processing of Aloe vera Gel Promotes Cytotoxicity on Human Intestinal Cells HT-29.

Aloe vera products, both in food and cosmetics, are becoming increasingly popular due to their claimed beneficial effects, which are mainly attributed to the active compound acemannan. Usually, these end products are based on powdered starting materials. High temperatures during the drying process to obtain the starting materials have several advantages, like shortening the drying time, eliminating toxic aloin and reducing bacterial contamination. Nevertheless, there are two major drawbacks: first, at temperatures of 80 °C or higher, structural changes in acemannan, especially its deacetylation (>46%), are triggered, which does not happen at lower temperatures (14% at 60 °C); secondly, a toxic principle is formed at higher temperatures, resulting in a higher cytotoxicity. Thus, two temperature-dependent but opposing effects cause with a median cytotoxic concentration of CC50 = 0.4× a peak of cytotoxicity at 80 °C; at 60 °C this cytotoxic substance is not formed and at 100 °C aloin is more readily eliminated, resulting in a CC50 = 1.1× and CC50 = 1.4×, respectively. The cytotoxic substance generated by dry heat at 80 °C is not a modified polysaccharide because its polysaccharide-enriched alcohol-insoluble fraction is with CC50 = 0.9× less cytotoxic. Moreover, this substance is polar enough to be washed away with ethanol. Additionally, when Aloe gel is heated at 80 °C under humid conditions (pasteurization), the cytotoxicity does not increase (CC50 = 1.6×). Finally, to produce powdered starting materials from Aloe gel, it is recommended to use temperatures of around 60 °C in order to preserve the acemannan structure (and thus biological activity) and the low cytotoxicity.

A Ferrofluid with Surface Modified Nanoparticles for Magnetic Hyperthermia and High ROS Production.

A ferrofluid with 1,2-Benzenediol-coated iron oxide nanoparticles was synthesized and physicochemically analyzed. This colloidal system was prepared following the typical co-precipitation method, and superparamagnetic nanoparticles of 13.5 nm average diameter, 34 emu/g of magnetic saturation, and 285 K of blocking temperature were obtained. Additionally, the zeta potential showed a suitable colloidal stability for cancer therapy assays and the magneto-calorimetric trails determined a high power absorption density. In addition, the oxidative capability of the ferrofluid was corroborated by performing the Fenton reaction with methylene blue (MB) dissolved in water, where the ferrofluid was suitable for producing reactive oxygen species (ROS), and surprisingly a strong degradation of MB was also observed when it was combined with H2O2. The intracellular ROS production was qualitatively corroborated using the HT-29 human cell line, by detecting the fluorescent rise induced in 2,7-dichlorofluorescein diacetate. In other experiments, cell metabolic activity was measured, and no toxicity was observed, even with concentrations of up to 4 mg/mL of magnetic nanoparticles (MNPs). When the cells were treated with magnetic hyperthermia, 80% of cells were dead at 43 °C using 3 mg/mL of MNPs and applying a magnetic field of 530 kHz with 20 kA/m amplitude.

Green Metallic Nanoparticles for Cancer Therapy: Evaluation Models and Cancer Applications.

Metal-based nanoparticles are widely used to deliver bioactive molecules and drugs to improve cancer therapy. Several research works have highlighted the synthesis of gold and silver nanoparticles by green chemistry, using biological entities to minimize the use of solvents and control their physicochemical and biological properties. Recent advances in evaluating the anticancer effect of green biogenic Au and Ag nanoparticles are mainly focused on the use of conventional 2D cell culture and in vivo murine models that allow determination of the half-maximal inhibitory concentration, a critical parameter to move forward clinical trials. However, the interaction between nanoparticles and the tumor microenvironment is not yet fully understood. Therefore, it is necessary to develop more human-like evaluation models or to improve the existing ones for a better understanding of the molecular bases of cancer. This review provides recent advances in biosynthesized Au and Ag nanoparticles for seven of the most common and relevant cancers and their biological assessment. In addition, it provides a general idea of the in silico, in vitro, ex vivo, and in vivo models used for the anticancer evaluation of green biogenic metal-based nanoparticles.

Sterilized chitosan-based composite hydrogels: Physicochemical characterization and in vitro cytotoxicity.

Gelatin/chitosan/polyvinyl alcohol hydrogels were fabricated at different polymer ratios using the freeze-drying and sterilized by steam sterilization. The thermal stability, chemical structure, morphology, surface area, mechanical properties, and biocompatibility of hydrogels were evaluated by simultaneous thermal analysis, Fourier transform infrared spectroscopy, X-ray diffraction, confocal microscopy, adsorption/desorption of nitrogen, rheometry, and 3-4,[5-dimethylthiazol-2-yl]-2,5-diphenyl tetrazolium bromide cell viability assay (MTT assay), respectively. The samples showed a decomposition onset temperature below 253.3 ± 4.8°C, a semicrystalline nature, and a highly porous structure. Hydrogels reached the maximum water uptake in phosphate-buffered saline after 80 min, showing values from nine to twelve times their dry mass. Also, hydrogels exhibiting a solid-like behavior ranging from 2,567 ± 467 to 48,705 ± 2,453 Pa at 0.1 rad/s (low frequency). The sterilized hydrogels showed low cytotoxicity (cell viability > 70%) to the HT29-MTX-E12 cell line. Sterilized hydrogels by steam sterilization can be good candidates as scaffolds for tissue engineering applications.

In Vitro Immunomodulatory Effect of Food Supplement from Aloe vera.

Food industries typically use Aloe vera as concentrated (100× to 200×) and dried powders in their final products. These powders are obtained by extrusion of Aloe inner leaf gel (ILG) or Aloe whole leaf (WLP); the juice is filtered through diatomaceous earth and activated carbon before spray drying at temperatures below 70 °C. In another process, Aloe inner leaf gel was dried at ~80 °C and mashed to a powder rich in high molecular weight fibres and soluble polysaccharides (ILF). In contrast to ILG and WLP, the ILF sample was cytotoxic for the human intestinal cell line Caco-2 (CC50 = 1 g/l), even at concentrations below the recommended dose for human consumption. At lower concentrations (250 mg/l) with LPS challenged macrophage-like THP-1 cells decreased by 40% the release of the anti-inflammatory cytokine IL-10, whereas the release of the proinflammatory cytokine IL-1ß increased by 35% (compared to untreated but challenged macrophage-like THP-1 cells). Unexpectedly, under the same conditions, the less cytotoxic ILG and WLP, both samples with a lower fibre content, significantly increased (up to 2.4 times) the release of IL-10, while the concentration of IL-1ß remained unaltered and of TNFα decreased by 35%. Even more interesting is that a treatment of the ILF sample with activated carbon reduced its cytotoxicity and increased the IL-10 release (3.1 times). Based on these results, we suggest applying an activated carbon treatment on Aloe-starting products, which have high fibre content and have received high temperature treatment, in order to reduce their cytotoxicity and improve their immunomodulatory properties.

In Vitro Bioactivity of Methanolic Extracts from Amphipterygium adstringens (Schltdl.) Schiede ex Standl., Chenopodium ambrosioides L., Cirsium mexicanum DC., Eryngium carlinae F. Delaroche, and Pithecellobium dulce (Roxb.) Benth. Used in Traditional Medicine in Mexico.

Seven out of eight methanolic extracts from five plants native to Mexico were inactive against ten bacterial strains of clinical interest. The fruit extract of Chenopodium ambrosioides inhibited the bacteria Enterococcus faecalis (MIC = 4375 µg/ml), Escherichia coli (MIC = 1094 µg/ml), and Salmonella typhimurium (MIC = 137 µg/ml). The fruit extract of C. ambrosioides was with CC50 = 45 µg/ml most cytotoxic against the cell-line Caco-2, followed by the leaf extract from Pithecellobium dulce (CC50 = 126 µg/ml); interestingly, leaves of C. ambrosioides (CC50 = 563 µg/ml) and bark of P. dulce (CC50 = 347 µg/ml) extracts were much less cytotoxic. We describe for the first time the cytotoxic effect from extracts of the aerial parts and the flowers of Cirsium mexicanum (CC50 = 323 µg/ml and CC50 = 250 µg/ml, resp.). Phytochemical analysis demonstrated for both extracts high tannin and saponin and low flavonoid content, while terpenoids were found in the flowers. For the first time we report a cytotoxicological study on an extract of Eryngium carlinae (CC50 = 356 µg/ml) and likewise the bark extract from Amphipterygium adstringens (CC50 = 342 µg/ml). In conclusion the fruit extract of C. ambrosioides is a potential candidate for further biological studies.

Are biological control agents, isolated from tropical fruits, harmless to potential consumers?

Postharvest losses of fruits and vegetables can reach up to 25% in developed and up to 50% in developing countries. (Sub)tropical fruits are especially susceptible because their protecting peel can easily be damaged. Traditionally used pesticides are associated to environmental pollution and possible harmful health effects. An alternative are biocontrol agents (BCA), means bacteria or yeasts applied onto the fruits to inhibit the growth of phytopathogens. Many reports on their effectiveness have been published, however, reports on their harmlessness to consumers are still rare. Culture extracts of six BCAs, tested on two human lines (Caco-2, HeLa), exhibited no cytotoxic effect, when used directly (1×) to protect the fruits; however, when they are 5×overconcentrated, the confluence of proliferating cells was reduced, but not of differentiated Caco-2. In both cases necrosis was not increased. On proliferating cells, the 5×-extract from Cryptococcus laurentii or Debaryomyces hansenii reduced lysosome functionality and the 6.25×extract from Meyerozyma guilliermondii or Candida famata increased membrane permeability, while only the 25×-extract from M. guilliermondii or M. caribbica reduced slightly the metabolic activity. The extract of Bacillus subtilis showed no cytotoxic effect up to 10× concentration. Overall, their low cytotoxicity combined with high biodegradability make these products suitable for sustainable agriculture.

Supervisor-subordinate age dissimilarity and performance ratings: the buffering effects of supervisory relationship and practice.

Using 394 pairs of employees and their immediate supervisors working in the Information and Communication Technology (ICT) sector in three northern European countries, this study examined the effect of workplace moderators on the link between relational demography and supervisor ratings of performance. Directional age differences between superior and subordinate (i.e., status incongruence caused when the supervisor is older or younger than his/her subordinate) and non-directional age differences were used as predictors of supervisor ratings of occupational expertise. The quality of the supervisor-subordinate relationship and the existence of positive age-related supervisory practices were examined as moderators of this relationship. The results provide no support for a relationship between directional age differences and age-related stereotyping by supervisors in ratings of performance, neither for the effects of age-related supervisory practices. However, high quality supervisor-subordinate relationships did moderate the effects of age dissimilarity on supervisory ratings. The implications of these findings for performance appraisal methodologies and recommendations for further research are discussed.

Shift work and cancer: the evidence and the challenge.

BACKGROUND: In 2007, the International Agency for Research on Cancer (IARC) classified shift work with circadian disruption or chronodisruption as a probable human carcinogen. Short-term disturbances of biological 24-hour-rhythms following exposures to light and darkness at unusual times are well-known as "jet-lag" and "shift-lag" symptoms. However, that chronic disturbances or disruptions of timely sequenced circadian rhythms (chronodisruption) should contribute to long-term developments of cancer is a relatively new concept. This review provides background and practical information with regard to the open question "does shift-work cause cancer?" METHODS: Overview on the basis of a selective literature search via Medline and ISI Web of Knowledge until 2009 from the viewpoints of occupational medicine, epidemiology, chronobiology, and occupational science. RESULTS: The postulated causal links between shift-work and cancer in humans are biologically plausible in the light of experimental findings, but to date we lack epidemiological studies which could describe or exonerate risks in humans. Monetary compensation has already been paid for such cases in at least one country (Denmark). In Germany, however, according to the applicable law, a new occupational disease can only be recognized when certain conditions for the recognition of "general scientific merit" have been met. We present the current state of knowledge regarding prevention. CONCLUSION: While causal links between shift-work and cancer developments are not established, future shift-work planning should pay more attention to insights from occupational medicine, chronobiology, and occupational science.

Is health, measured by work ability index, affected by 12-hour rotating shift schedules?

Two forms of continuously forward rotating 12-h shift schedules exist at BASF's Ludwigshafen site. These shift schedules were compared with a daytime working system to investigate potential differential effects on employee's health status assessed with the Work Ability Index (WAI). In the 3 x 12 system, a 12-h day shift is followed 24 h later by a 12-h night shift, and after a day off the employee returns to the day shift. The 4 x 12 schedule follows the same pattern except that there are 2 days off between the night and next day shift. A total of 924 participants (278 3 x 12 and 321 4 x 12 shiftworkers and 325 day workers) were recruited. A self-administered questionnaire was used to obtain information about shiftwork schedule, demographic characteristics, and lifestyle and social factors, and the WAI was applied. The outcomes of interest were the WAI sum score and its seven dimensions. In examining the relationship with the WAI categories, a Proportional Odds Model (POM) was used to identify the potential determinants. Logistic regression models were used to estimate the impact of age on single dimensions of WAI after adjustment for potential confounding factors. Increasing age and obesity (BMI > or = 30) were the only significant determinants of poorer WAI. Although a positive association was found linking the second WAI dimension (work ability in relation to job demands) with age, an inverse association was demonstrated consistently between age and the third and fourth WAI dimensions, i.e., number of diagnosed diseases and estimated work impairment due to disease, after adjustment for potential confounders. The age-dependency was moderate overall, but seemed to be stronger among shift- than day workers, although this difference did not reach statistical significance. There was no significant differential impact of the working time systems on the WAI sum score or on the individual WAI dimensions. Thus, there is no indication of an excessive adverse health impact of these shift schedules compared to day work, to the extent that health can be measured by the WAI.

Shift work, chronodisruption and cancer?--The IARC 2007 challenge for research and prevention and 10 theses from the Cologne Colloquium 2008.

In October of 2007, an IARC panel of 24 scientists systematically evaluated epidemiologic, experimental, and mechanistic data and concluded that shift work that involves circadian or chronodisruption is probably carcinogenic in humans. In view of the possible scope of the problem--shift work is widespread and unavoidable on one hand and breast cancer and prostate cancer, which may be causally associated with chronodisruption, are epidemic worldwide on the other--German representatives of science and occupational medicine discussed the experimental and epidemiologic background and possible implications of the challenge identified by the International Agency for Research on Cancer (IARC) at a colloquium in Cologne in September 2008. This overview summarizes the key ideas presented at the Cologne Colloquium and offers 10 theses concerning the need for targeted studies and the necessity to develop possible means of prevention.

Numb3 is an endocytosis adaptor for the inflammatory marker P-selectin.

The endocytic protein Numb3 was found to bind to the cytosolic tail of the leukocyte adhesion receptor P-selectin. The N-terminal phosphotyrosine-binding (PTB) domain of Numb3 is responsible for this activity. An alanine scan revealed the FTNAAFD sequence as recognition region in P-selectin. Structural modeling of the interaction between the Numb PTB domain and the P-selectin tail suggests that both phenylalanines within the recognition sequence fit into hydrophobic cavities of the PTB surface. Their exchange for alanine gave Numb-negative mutants detaining the inhibition of P-selectin endocytosis by Numb PTB overexpression. Cells stable expressing P-selectins internalized the negative mutants markedly slower than the wild type. Consistent with other reports on the phosphorylation of Numb, we found that only the dephospho-Numb is able to bind P-selectin. Our observations demonstrate that Numb3 is an endocytic receptor for P-selectin and may be responsible for the rapid internalization of P-selectin when endothelial activation ends.

Extended work periods.

A literature review of 105 studies on the effects of extended daily working hours was conducted. Potential negative effects of extended working hours are discussed: More accidents on the job; more accidents off the job; reduced duration and quality of sleep due to moonlighting; sleepiness; reduced alertness; fatigue; adverse effects on performance; prolonged toxic exposure; adverse effects on health; increased absenteeism; problems communicating with managers; and problems while driving home. Potential positive effects of extended working hours are discussed: Less travel time and costs; more time for the family, social life, and domestic duties; increased satisfaction with working hours; fewer handovers; and less overtime. No firm conclusions can be drawn because of the partly contradictory results and the methodological problems of many studies. However, caution is advised when considering the introduction of extended work shifts, particularly where public safety is at stake. A checklist is provided (concerning work load, breaks, staffing level, systematic assessments of health and safety factors) to support decisions for or against the use of extended work shifts.

Compensation for unfavorable characteristics of irregular individual shift rotas.

Some employees of TV companies, such as those who produce remote TV programs, have to cope with very irregular rotas and many short-term schedule deviations. Many of these employees complain about the negative effects of such on their wellbeing and private life. Therefore, a working group of employers, council representatives, and researchers developed a so-called bonus system. Based on the criteria of the BESIAK system, the following list of criteria for the ergonomic assessment of irregular shift systems was developed: proportion of night hours worked between 22 : 00 and 01 : 00 h and between 06 : 00 and 07 : 00 h, proportion of night hours worked between 01 : 00 and 06 : 00 h, number of successive night shifts, number of successive working days, number of shifts longer than 9 h, proportion of phase advances, off hours on weekends, work hours between 17 : 00 and 23 : 00 h from Monday to Friday, number of working days with leisure time at remote places, and sudden deviations from the planned shift rota. Each individual rota was evaluated in retrospect. If pre-defined thresholds of criteria were surpassed, bonus points were added to the worker's account. In general, more bonus points add up to more free time. Only in particular cases was monetary compensation possible for some criteria. The bonus point system, which was implemented in the year 2002 for about 850 employees of the TV company, has the advantages of more transparency concerning the unfavorable characteristics of working-time arrangements, incentive for superiors to design "good" rosters that avoid the bonus point thresholds (to reduce costs), positive short-term effects on the employee social life, and expected positive long-term effects on the employee health. In general, the most promising approach to cope with the problems of shift workers in irregular and flexible shift systems seems to be to increase their influence on the arrangement of working times. If this is not possible, bonus point systems may help to achieve greater transparency and fairness in the distribution of unfavorable working-time arrangements within a team, and even reduce the unnecessary unfavorable aspects of shift systems.

Sorting nexin 17, a non-self-assembling and a PtdIns(3)P high class affinity protein, interacts with the cerebral cavernous malformation related protein KRIT1.

The mammalian sorting nexin (SNX) proteins are involved in the endocytosis and the sorting machinery of transmembrane proteins. Additionally to the family defining phox homology (PX) domain, SNX17 is the only member with a truncated FERM (4.1, ezrin, radixin, and moesin) domain and a unique C-terminal region (together designated as FC unit). By gel filtration and lipid overlay assays we show that SNX17 is a non-self-assembling and a PtdIns(3)P high class affinity protein. A SNX17 affinity to any other phosphoinositides was not detected. By yeast two-hybrid- and GST-trapping assays we identified KRIT1 (krev1 interaction trapped 1) as a new specific interaction partner of the FC unit of SNX17. KRIT1 binds SNX17 by its N-terminal region like the known interaction partner ICAP1alpha (integrin cytoplasmic domain-associated protein-1). The interaction was also detected in HEK 293 cells transiently expressing GFP-tagged KRIT1 and Xpress-tagged SNX17. KRIT1 mutations cause cerebral cavernous malformation (CCM1). Our finding suggests a SNX17 involvement in the indicated KRIT1 function in cell adhesion processes by integrin signaling.

Functions of sorting nexin 17 domains and recognition motif for P-selectin trafficking.

SNX17 is a member of the sorting nexin family (SNX), a group of hydrophilic proteins whose common characteristic property is a phox homology (PX) domain. The PX domain directs SNXs to phosphatidylinositides containing membranes of the endosomal compartment, where the SNXs are involved in the sorting of transmembrane proteins. SNX17 is known to interact with P-selectin and the LDL receptor family. Here, we report that the PX domain of SNX17 specifically binds to phosphatidylinositol 3-phosphate-containing membranes. The functional part of SNX17 that binds P-selectin or Patched (PTCH) consists of a truncated FERM domain and a unique C terminus together (FC-unit). In a yeast two-hybrid analysis a putative recognition motif for the FC-unit was revealed within P-selectin as FxNaa(F/Y). When HepG2 cells overexpress P-selectin together with SNX17, SNX17 changes its distribution from early endosomes to lysobisphosphatidic acid-containing late endosomes. Furthermore, overexpressed SNX17 restrains P-selectin in the outer membrane of the late endosomal compartment, thus preventing the normal lysosomal accumulation of P-selectin. These results suggest that the PX domain is necessary for the intracellular localisation, while the FC-unit is required for cargo recognition. We hypothesise that the expression level of SNX17 may regulate the lysosomal degradation, at least for P-selectin, by suppressing its entry into the inner vesicles of the multi-vesicular bodies (MVBs).

Sorting nexin 17 accelerates internalization yet retards degradation of P-selectin.

The transient appearance of P-selectin on the surface of endothelial cells helps recruit leukocytes into sites of inflammation. The tight control of cell surface P-selectin on these cells depends on regulated exocytosis of Weibel-Palade bodies where the protein is stored and on its rapid endocytosis. After endocytosis, P-selectin is either sorted via endosomes and the Golgi apparatus for storage in Weibel-Palade bodies or targeted to lysosomes for degradation. A potential player in this complex endocytic itinerary is SNX17, a member of the sorting nexin family, which has been shown in a yeast two-hybrid assay to bind P-selectin. Here, we show that overexpression of SNX17 in mammalian cells can influence two key steps in the endocytic trafficking of P-selectin. First, it promotes the endocytosis of P-selectin from the plasma membrane. Second, it inhibits the movement of P-selectin into lysosomes, thereby reducing its degradation.

Preventive and compensatory measures for shift workers.

Shift systems are known to be associated with a variety of psychosocial and physiological problems that can affect the health of workers. This review focuses on measures that can be taken to optimize the well-being of shift workers and to identify ill-health at an early stage. The discussion includes specific aspects of the design of shift systems, taking account of variation in the views and circumstances of employees, and strategies to combat sleepiness at work and elsewhere. Although an ideal shift system does not exist, a wholistic approach comprising education of managers, employees and their families can ameliorate some of the health consequences.