Wortmannin induces MCF-7 breast cancer cell death via the apoptotic pathway, involving chromatin condensation, generation of reactive oxygen species, and membrane blebbing.

BACKGROUND: Apoptosis can be used as a reliable marker for evaluating potential chemotherapeutic agents. Because wortmannin is a microbial steroidal metabolite, it specifically inhibits the phosphatidyl inositol 3-kinase pathway, and could be used as a promising apoptosis-based therapeutic agent in the treatment of cancer. The objective of this study was to investigate the biomolecular mechanisms involved in wortmannin-induced cell death of breast cancer-derived MCF-7 cells. METHODS AND RESULTS: Our experimental results demonstrate that wortmannin has strong apoptotic effects through a combination of different actions, including reduction of cell viability in a dose-dependent manner, inhibition of proliferation, and enhanced generation of intracellular reactive oxygen species. CONCLUSION: Our findings suggest that wortmannin induces MCF-7 cell death via a programmed pathway showing chromatin condensation, nuclear fragmentation, reactive oxygen species, and membrane blebbing, which are characteristics typical of apoptosis.

Exoskeletal thinning in Cephalotes atratus ants (Hymenoptera: Formicidae) parasitized by Myrmeconema neotropicum (Nematoda: Tetradonematidae).

Some parasites modify the color of their arthropod hosts, presumably to facilitate transmission to a new host. Mechanisms for such changes often are unknown, but altered exoskeletal color in adult insects typically occurs via structural modifications or redistribution of pigments. Here, we examine the cuticle structure of workers of the Neotropical canopy ant Cephalotes atratus infected with the nematode Myrmeconema neotropicum. We hypothesized that the conspicuous red color of the gaster (the globular posterior body region) of infected ants results from structural changes, specifically localized exoskeletal thinning. We used scanning electron microscopy to quantify the thickness of gaster cuticle in healthy and infected ants. For comparison, we also measured the cuticle thickness of the head of each ant, which is black in both infected and healthy individuals. The gaster cuticle was 23% thinner in infected ants (average ±SE: 14.8 ± 1.02 µm) versus healthy ants (19.2 ± 0.65 µm) after correcting for body size. In contrast, the thickness of the head exoskeleton was similar among groups. We conclude that parasite-induced thinning of the exoskeleton is associated with the red color of the gaster. Other mechanisms, including translocation or leaching of melanin (by the ant or the parasite, respectively) may operate in concert with thinning to effect the color change, and would be an appropriate extension of this research.

Nickel nanowires induced and reactive oxygen species mediated apoptosis in human pancreatic adenocarcinoma cells.

BACKGROUND: The ability to evade apoptosis is one of the key properties of cancer. The apoptogenic effect of nickel nanowires (Ni NWs) on cancer cell lines has never been adequately addressed. Due to the unique physicochemical characteristics of Ni NWs, we envision the development of a novel anticancer therapeutics specifically for pancreatic cancer. Thus, we investigated whether Ni NWs induce ROS-mediated apoptosis in human pancreatic adenocarcinoma (Panc-1) cells. METHODS: In this study Ni NWs were fabricated using the electrodeposition method. Synthesized Ni NWs were physically characterized by energy dispersive X-ray analysis, UV-Vis spectroscopy of NanoDrop 2000 (UV-Vis), magnetization study, scanning electron microscopy, and transmission electron microscopy. Assessment of morphological apoptotic characteristics by phase contrast microscopy (PCM), Ni-NWs-induced apoptosis staining with ethidium bromide (EB) and acridine orange (AO) followed by fluorescence microscopy (FM) was performed. For molecular biological and biochemical characterization, Panc-1 cell culture and cytotoxic effect of Ni NWs were determined by using 3-(4, 5-dimethylthiazol-2-yl)-2, 5-diphenyl tetrazolium bromide (MTT) assay. Quantitative apoptosis was analyzed by flow cytometry staining with propidium iodide through cell cycle arrest and generation of ROS using 2', 7'-dichlorofluorescein diacetate fluorescence intensity. In all experiments, Panc-1 cancer cells without any treatment were used as the negative controls. RESULTS: The intracellular uptake of Ni NWs through endocytosis by Panc-1 cells was observed by PCM. EB and AO staining of FM and MTT assay qualitatively and quantitatively confirmed the extent of apoptosis. Flow cytometric cell cycle arrest and ROS generation indicated Ni NWs as inducers of apoptotic cell death. CONCLUSION: We investigated the role of Ni NWs as inducers of ROS-mediated apoptosis in Panc-1 cells. These results suggested that Ni NWs could be an effective apoptotic agent for Panc-1 cells and have good potential for further research into a clinical treatment selective for pancreatic cancer.

A new mermithid species (Nematoda) from chironomid imagos eclosing from Lake Ozawindib in northern Minnesota.

Hydromermis biesboeri n. sp. (Nematoda: Mermithidae) is described from the hemocoel of imagos of chironomid host species eclosing from Lake Ozawindib in northern Minnesota in July and August 2008 and 2009. The new species is distinguished from the other 28 described species in the genus by terminal mouth, wide oval (male) and long oval (female) amphids with a dorsal commissure, tapered and rounded posterior ends, split base of the spicule, short protractor muscles in males, and by females having an S-shaped vagina with equal-length terminal limbs. The new species represents the 10th Hydromermis mermithid species described, or reported, from the region and the second from Lake Ozawindib.

A new genus and species of mermithid (Nematoda) emerging from chironomid imagos eclosing from the Mississippi River headwaters area in northern Minnesota.

Fibromermis roarki n. gen., n. sp. (Nematoda: Mermithidae) is described from the hemocoel of chironomid imagos of Ablabesmyia monilis (L.) eclosing from the headwaters of the Mississippi River at Gulsvig Landing in northern Minnesota in the summer of 2007. The new genus is described, distinguished from the most closely related tetrapapillate aquatic mermithid genus, and the new species described. Intensity and sex specificity of infection are recorded from 28 host adult chironomids of a single cohort. The new species represents the fourth tetrapapillate mermithid species described from the area and the third tetrapapillate genus.

A life-span extending form of autophagy employs the vacuole-vacuole fusion machinery.

While autophagy is believed to be beneficial for life-span extension, it is controversial which forms or aspects of autophagy are responsible for this effect. We addressed this topic by analyzing the life span of yeast autophagy mutants under caloric restriction, a longevity manipulation. Surprisingly, we discovered that the majority of proteins involved in macroautophagy and several forms of microautophagy were dispensable for life-span extension. The only autophagy protein that is critical for life-span extension was Atg15, a lipase that is located in the endoplasmic reticulum (ER) and transported to vacuoles for disintegrating membranes of autophagic bodies. We further found that vacuole-vacuole fusion was required for life-span extension, which was indicated by the shortened life span of mutants missing proteins (ypt7Delta, nyv1Delta, vac8Delta) or lipids (erg6Delta) involved in fusion. Since a known function of vacuole-vacuole fusion is the maintenance of the vacuole membrane integrity, we analyzed aged vacuoles and discovered that aged cells had altered vacuolar morphology and accumulated autophagic bodies, suggesting that certain forms of autophagy do contribute to longevity. Like aged cells, erg6Delta accumulated autophagic bodies, which is likely caused by a defect in lipase instead of proteases due to the existence of multiple vacuolar proteases. Since macroautophagy is not blocked by erg6Delta, we propose that a new form of autophagy transports Atg15 via the fusion of vacuoles with vesicles derived from ER, and we designate this putative form of autophagy as secretophagy. Pending future biochemical studies, the concept of secretophagy may provide a mechanism for autophagy in life-span extension.

A new species of mermithid (Nematoda) in chironomids eclosing from a northern Minnesota bog.

Hydromermis haggardi n. sp. (Nematoda: Mermithidae) is described from the hemocoel of chironomid imagos eclosing from Lake Alice Bog in northern Minnesota in June and July 2007. The species is distinguished from the other 27 described members of the genus by the subventral mouth, tulip shape to anterior end of esophagus, colorless vesicular trophosome, single nonbifurcated spicule, short body lengths of both males and females, structure of the S-shaped vagina, decrease in body width at vulva, brevity of postparasitic stage, and the greater development of a posterior ventral protractor muscle in the males. An anomalous adult male bearing large oval structures in the posterior testis is reported. Evidence is presented for the developmental interaction of the new species with Lanceimermis palustris Johnson and Kleve, 2004.

A new mermithid nematode from Ablabesmyia monilis (L.) (Diptera: Chironomidae) in northern Minnesota.

Limnomermis steineri n. sp. (Nematoda: Mermithidae) is described from the hemocoel of male, female, and intersex imagos of Ablabesmyia (s. str.) monilis (L.) eclosing from Lake Ozawindib and Lake Alice Bog in northern Minnesota during summers 2000-2005. The species is distinguished from other described members of the genus. Paramermis rosea is returned to Limnomermis. A review of adequately described members of the genus is included. One species of the genus has been reported previously from the Nearctic Realm. Emphasis is placed on the need for host identification, larger number of specimens of both mermithid sexes, and information on the intensity of infections to enable more precise range comparisons in mermithid biometrics.

Hydromermis osami n. sp. (Nematoda: Mermithidae) from chironomids eclosing from northern Minnesota waterways.

Hydromermis osami n. sp. (Nematoda: Mermithidae) is described from the hemocoel of 2 species of Rheotanytarsus chironomid imagos eclosing from Lake Ozawindib and Gulsvig Landing in northern Minnesota during August 2002 and June and July 2006. The species is distinguished from the other 26 described members of the genus by the terminal mouth; oval, opaque, thick walled amphids, wider than long; acute posterior end; single spicule not bifurcated proximally; well-developed uterine and vulval limbs of the S-shaped vagina; absence of bursal sleeve; absence of nutrient vesicles in the trophosome; and an esophagus length over 40% of body length. Members of the new species emerge from the hosts as adults. The other described Hydromermis species, number of specimens, location, and known hosts are tabulated.

Lanceimermis palustris n. sp. and Telomermis palustris n. sp. (Nematoda: Mermithidae) parasites of chironomid (Insecta: Diptera) adults from Lake Alice Bog in northern Minnesota.

In the summer of 2003, chironomid imagoes eclosing from Lake Alice Bog in Minnesota yielded mermithid nematodes of 2 new species. Lanceimermis palustris n. sp. is distinguished from the other 14 species of the genus by the subventral mouth position, thinness of the hook-shaped spicule, body index, and structure of the male tail muscles. Telomermis palustris n. sp. is distinguished from the only other species of the genus by the structure of the amphids, body length, maximum body width, length of the esophagus, and the presence of a minute larval terminal horn. Lanceimermis palustris n. sp. hosts were Chironomus maturus Johannsen and T. palustris n. sp. hosts were Tanytarsus mendax Kieffer, Paratanytarsus nr. dissimilis n. sp., and Micropsectra polita (Malloch). In both mermithid species, the percent of mermithid males per host increased with the intensity of the infection.

Strelkovimermis papillosus n. sp. (Nematoda: Mermithidae), a parasite of chironomid (Insecta: Diptera) adults from the headwaters of the Mississippi River in northern Minnesota.

On 18 August 2002, chironomid imagoes of Rheotanytarsus sp. emerged from the upper Mississippi River in Minnesota and yielded distinctive mermithid nematodes of a new mermithid species. Strelkovimermis papillosus n. sp. is distinguished from the other 14 species of the genus by the presence of unusually large cephalic papillae encircling the mouth and forming a rosette with the mouth in the center and by the absence of a fixator muscle in the males. Additionally, both sexes have very acute posterior ends, long amphids, and a long stoma. Strelkovimermis is revised by eliminating nondiscriminating parameters and accommodating the 15 known species. Intrageneric characteristics useful in separating species of Strelkovimermis are listed. Intensity of infection and intensity of infection versus sex were determined from 41 hosts. Where known, the hosts and geographical distribution are given for all 15 Strelkovimermis species.

Strelkovimermis rubtsovi n. sp. and Strelkovimermis ozawindibi n. sp. (Nematoda: Mermithidae) parasitizing chironomid (Insecta: Diptera) adults eclosing from northern Minnesota glacial lakes.

Two new species of Strelkovimermis are described from chironomid imagoes eclosing from northern Minnesota glacial lakes. The 2 species are distinguished from the other 12 species in the genus by terminal mouths, rounded or nippled posterior ends, short buccal funnels, short terminal limbs of the S-shaped vagina, and presence of a bursal sleeve. Strelkovimermis rubtsovi n. sp. is distinguished from S. ozawindibi n. sp. by the presence of a dorsal protractor. Procladius (Psilotanypus) bellus (Loew) is the host of S. rubtsovi. The chironomid host of S. ozawindibi has not been determined. An artificial key is provided to distinguish the 14 species of the genus.