Heavy metal bioaccumulation in honey bee matrix, an indicator to assess the contamination level in terrestrial environments.

The most significant risk factor for organisms living in an environment contaminated by heavy metals is the metal bioavailability. Therefore, an efficient ecotoxicological approach to metal contamination is the measure of bioaccumulation level in target organisms. In this work, we characterized the heavy metal bioaccumulation in honey bees, Apis mellifera ligustica, collected at 35 sites from Umbria (Central Italy). The comparison of our data with selected Italian investigations revealed metal bioaccumulation in honey bee matrix of the same order of magnitude, with Cd showing a higher variability. To generalize the results, we developed a Honeybee Contamination Index (HCI) based on metal bioaccumulation in honey bees. An application of the HCI to the present dataset revealed cases of low (sixteen sites), intermediate (eighteen sites), and high (one site) metal contaminations. The comparison of HCI values from the Umbrian dataset with values calculated for other Italian and European metadata showed that most of the Umbrian sites fell in the portion of low and intermediate contamination conditions. HCI represented a reliable tool that provided a piece of concise information on metal contamination in terrestrial environments. Parallel to this effort, we have determined, the metal concentrations in the airborne particulate matter (PM10) at three regional background-monitoring stations in Umbria. These stations are representative of the average air quality of the areas of the investigated apiaries. A comparative analysis of metal enrichment factors in PM10, and honey bees suggested that the contamination in the bees was related to the PM10 values only to a minor extent. On the other side, a clear enrichment of metals such as Cd, Mn, Zn, and Cu in the honey bees appeared to depend on very local conditions and was probably related to the use of pesticides and fertilizers, and the resuspension of the locally contaminated soils and agriculture residues.

Disruption of the anaphase-promoting complex confers resistance to TTK inhibitors in triple-negative breast cancer.

TTK protein kinase (TTK), also known as Monopolar spindle 1 (MPS1), is a key regulator of the spindle assembly checkpoint (SAC), which functions to maintain genomic integrity. TTK has emerged as a promising therapeutic target in human cancers, including triple-negative breast cancer (TNBC). Several TTK inhibitors (TTKis) are being evaluated in clinical trials, and an understanding of the mechanisms mediating TTKi sensitivity and resistance could inform the successful development of this class of agents. We evaluated the cellular effects of the potent clinical TTKi CFI-402257 in TNBC models. CFI-402257 induced apoptosis and potentiated aneuploidy in TNBC lines by accelerating progression through mitosis and inducing mitotic segregation errors. We used genome-wide CRISPR/Cas9 screens in multiple TNBC cell lines to identify mechanisms of resistance to CFI-402257. Our functional genomic screens identified members of the anaphase-promoting complex/cyclosome (APC/C) complex, which promotes mitotic progression following inactivation of the SAC. Several screen candidates were validated to confer resistance to CFI-402257 and other TTKis using CRISPR/Cas9 and siRNA methods. These findings extend the observation that impairment of the APC/C enables cells to tolerate genomic instability caused by SAC inactivation, and support the notion that a measure of APC/C function could predict the response to TTK inhibition. Indeed, an APC/C gene expression signature is significantly associated with CFI-402257 response in breast and lung adenocarcinoma cell line panels. This expression signature, along with somatic alterations in genes involved in mitotic progression, represent potential biomarkers that could be evaluated in ongoing clinical trials of CFI-402257 or other TTKis.

Comparison of organochlorine pesticides, PCBs, and heavy metal contamination and of detoxifying response in tissues of Ameiurus melas from Corbara, Alviano, and Trasimeno Lakes, Italy.

Accumulation of cadmium (Cd), mercury (Hg), lead (Pb), chromium (Cr), hexachlorobenzene, gamma-HCH, DDTs, and PCBs has been investigated in the muscle of Ameiurus melas sampled during the same period from Lake Corbara, Alviano, and Trasimeno. Glutathione content and the enzymatic activities of glutathione reductase, glutathione peroxidase, catalase, glutathione S-transferase, and glyoxalase I were examined in gills, liver, and kidneys of each specimen. Catfish from Alviano, compared to those of Corbara and Trasimeno, showed the highest contamination of DDTs and PCBs and the lowest levels of biochemical parameters. Most likely, OCPs and PCBs content might be responsible for the compromised antioxidant status in these specimens.

N,S-bis-fluorenylmethoxycarbonylglutathione: a new, very potent inhibitor of mammalian glyoxalase II.

A very potent competitive inhibitor of mammalian glyoxalase II activity, N,S-bis-fluorenylmethoxycarbonylglutathione (DiFMOC-G) has been synthesized and characterized. The Ki value for inhibition of glyoxalase II purified from calf liver is 0.08 microM. The Ki values for glyoxalase I inhibitions range from 285 to 500 fold higher than the values obtained for glyoxalase II inhibitions, depending on the source of the enzyme. Among other enzymes involved in glutathione metabolism, such as glutathione S-transferase, glutathione reductase, and glutathione peroxidase, only glutathione S-transferase is inhibited to a small extent by DiFMOC-G. Diesters of DiFMOC-G were prepared in order to improve transport of DiFMOC-G into mammalian tumor cells (rat adrenal pheochromocytoma, PC-12) in culture. Among the diesters synthesized, diisopropyl DiFMOC-G was found to be the most inhibitory to cell viability, with a [I]0.5 value of 3 microM.

S-fluorenylmethoxycarbonyl glutathione and diesters: inhibition of mammalian glyoxalase II.

Inhibitors having high specificity toward mammalian glyoxalase II, but not glyoxalase I, were sought as part of a program to study glyoxalase enzyme function in mammalian cells. The compound, S-fluorenylmethoxycarbonyl glutathione (FMOC-G), was synthesized and found to be a competitive inhibitor of purified calf liver glyoxalase II (Ki = 2.1 mumol/l). Inhibition constants (Ki values) for the other glyoxalase enzyme, glyoxalase I, and the glutathione-requiring enzyme, glutathione S-transferase, from other sources, were found to be 17 and 25 mumol/l, respectively. FMOC-G is a very poor inhibitor of glutathione reductase and glutathione peroxidase. Diesters (dimethyl, diethyl, diisopropyl) of FMCO-G were also synthesized, as proinhibitors, to improve transport of FMOC-G into mammalian tumor cells (rat adrenal pheochromocytoma, PC-12) in culture. The diesters were inhibitory to cell growth and variability; the most effective of these, diisopropyl FMOC-G, exhibited an [I]0.5 value of approximately 275 mumol/l. Diesters of FMOC-G may be useful in studies of the glyoxalase enzyme system in cultured mammalian cells.