Anticancer activities of selected species of North American lichen extracts.

Cancer is the second leading cause of human deaths in the USA. Despite continuous efforts to treat cancer over the past 50 years, human mortality rates have not decreased significantly. Natural products, such as lichens, have been good sources of anticancer drugs. This study reports the cytotoxic activity of crude extracts of 17 lichen species against Burkitt's lymphoma (Raji) cells. Out of the 17 lichen species, extracts from 14 species showed cytotoxicity against Raji cells. On the basis of IC50 values, we selected Xanthoparmelia chlorochroa and Tuckermannopsis ciliaris to study the mechanism of cell death. Viability of normal lymphocytes was not affected by the extracts of X. chlorochroa and T. ciliaris. We found that extracts from both lichens decreased proliferation, accumulated cells at the G0 /G1 stage, and caused apoptosis in a dose-dependent manner. Both lichen extracts also caused upregulation of p53. The T. ciliaris extract upregulated the expression of TK1 but X. chlorochroa did not. We also found that usnic, salazinic, constictic, and norstictic acids were present in the extract of X. chlorochroa, whereas protolichesterinic acid in T. ciliaris extracts. Our data demonstrate that lichen extracts merit further research as a potential source of anticancer drugs.

DNA damage caused by inorganic particulate matter on Raji and HepG2 cell lines exposed to ultraviolet radiation.

Epidemiological studies have correlated exposure to ultraviolet-irradiated particulate matter with cardiovascular, respiratory, and lung diseases. This study investigated the DNA damage induced by two major inorganic particulate matter compounds found in diesel exhaust, ammonium nitrate and ammonium sulfate, on Burkitt's lymphoma (Raji) and hepatocellular carcinoma (HepG2) cell lines. We found a dose-dependent positive correlation of accumulated DNA damage at concentrations of ammonium nitrate (25 µg/ml, 50 µg/ml, 100 µg/ml, 200 µg/ml, 400 µg/ml) with ultraviolet exposure (250 J/m(2), 400 J/m(2), 600 J/m(2), 850 J/m(2)), as measured by the comet assay in both cell lines. There was a significant difference between the treated ammonium nitrate samples and negative control samples in Raji and HepG2 cells (p<0.001). Apoptosis was shown in Raji and HepG2 cells when exposed to high concentrations of ammonium nitrate (200 µg/ml and 400 µg/ml) for 1h in samples without ultraviolet exposure, as assessed by the comet assay. However, the level of apoptosis greatly diminished after ultraviolet exposure at these concentrations. Over a 24h period, at intervals of 1, 4, 8, 12, 18, and 24h, we also observed that ammonium nitrate decreased viability in Raji and HepG2 cell lines and inhibited cell growth. Ammonium sulfate-induced DNA damage was minimal in both cell lines, but there remained a significant difference (p<0.05) between the ultraviolet radiation treated and negative control samples. These results indicate that the inorganic particulate compound, ammonium nitrate, induced DNA strand breaks at all concentrations, and indications of apoptosis at high concentrations in Raji and HepG2 cells, with ultraviolet radiation preventing apoptosis at high concentrations. We hypothesize that ultraviolet radiation may inhibit an essential cellular mechanism, possibly involving p53, thereby explaining this phenomenon. Further studies are necessary to characterize the roles of apoptosis inhibition induced by DNA damage caused by inorganic particulate matter.

A Histopathology Study of Caspian Seal (Pusa caspica) (Phocidae, Mammalia) Liver Infected with Trematode, Pseudamphistomum truncatum (Rudolphi, 1819) (Opisthorchidae, Trematoda).

BACKGROUND: Main objective of this study was to investigate the invasive activity of the liver fluke, Pseudamphistomom truncatum against the Caspian seal (Pusa caspica) and was exemplified at the gross, light microscopy (LM) and electron microscopy (EM) levels. METHODS: The study was done on a freshly dead Caspian Seal in the southern coast of Caspian Sea. The checked Caspian seal probably being died of canine distemper virus and was found host to numerous parasites of four helminth species. RESULTS: P. truncatum caused edematous foci on the surface of the liver with prominent fluid accumulation. Sections of the liver viewed with LM had multiple necrotic areas with extensive hemorrhaging and disorganized hepatic lobules. Granulocytes and invasion of connective tissue were prominent. Whole worms were visible with invasive pathways through the host tissue. Damage to both hepatic ducts and blood vessels were prominent. At the EM level, organelles within the impacted hepatocytes were disorganized as exemplified by the cristae of the mitochondria and the endoplasmic reticulum. Parasite eggs were scattered throughout the tissue. CONCLUSION: It was shown that this trematode can be very pathogenic to Caspian Seal and as this only mammal of Caspian Sea is an endangered species; this needs more investigation toward control or possible treatment of this helminth.

The description and histopathology of Leptorhynchoides polycristatus n. sp. (Acanthocephala: Rhadinorhynchidae) from sturgeons, Acipenser spp. (Actinopterygii: Acipenseridae) in the Caspian Sea, Iran, with emendation of the generic diagnosis.

Of the three known species of Leptorhynchoides Kostylew 1924, two are reported from North American fishes: Leptorhynchoides aphredoderi Buckner and Buckner 1976 and Leptorhynchoides thecatus (Linton 1891) Kostylew 1924. The third species, Leptorhynchoides plagicephalus (Westrumb 1821) Kostylew 1924, is commonly found in the Caspian and Black Sea from at least four species of sturgeons including Acipenser stellatus Pallas 1771 and the Acipenser nudiventris Lovetzsky 1828 from which Leptorhynchoides polycristatus n. sp. was found. No taxonomic work has been reported for L. plagicephalus for the last 90 years. L. polycristatus n. sp. can be readily confused with L. plagicephalus because of many superficial similarities. Such similarities include the general shape of the trunk, proboscis, and organ systems. However, L. polycristatus is clearly distinguished from the other three species primarily by having (1) 19-20 proboscis hooks per row; (2) the shortest hooks are anterior and the longest at the middle; the opposite is true in L. plagicephalus; (3) with a cuticular collar enveloping the base of the proboscis hooks; (4) the surface of its proboscis hooks is ribbed; (5) with a broad collar of multiple rectangular cuticular crests encircling the anterior end of the trunk; this is the only member of Leptorhynchoides with such a structure; (6) with many large ovoid uninucleated cells in the subcuticular layer of the trunk; (7) with paired glandular clusters near the male reproductive opening and of suction cup-like sensory structures on the bursa; (8) with dorsoventral ligament across the vagina; (9) cement glands are in a cluster of eight arranged in two horizontal tiers of four glands each; (10) with female gonopore near terminal; (11) with structures interpreted as possible microtriches on the surface of the trunk; (12) and with thinner eggs. L. polycristatus caused extensive histopathological damage to host intestinal layers. The armed proboscis invades and attaches to the host mucosa causing villi compression and necrosis of the epithelial lining with subsequent hemorrhaging and granulocyte migration. No encapsulation of the acanthocephalan is visible, and the worm can migrate deep into the smooth muscle layers of the muscularis extrema. The presence of L. polycristatus in the lumen of the host intestine obstructs and damages the absorbing surface of the host affecting the nutritional potential. Dead, necrotic host epithelial tissue and remnants of villi and crypts are visible.

The description of Mediorhynchus africanus n. sp. (Acanthocephala: Gigantorhynchidae) from galliform birds in Africa.

Mediorhynchus africanus n. sp. is described from specimens collected from the helmeted guinea fowls, Numida meliagris Linn. 1758 in Kruger National Park and elsewhere in subSaharan Africa from the same and other galliform birds. These specimens were previously assigned to Mediorhynchus gallinarum Bhaleroa (Proc Zool Soc Lond Ser B Syst Morph 107:199-203, 1937) described from chickens, Gallus gallus L. in India and subsequently reported from other Asian countries. The identification of the African forms as M. gallinarum was based on similarities in the structure and measurements of the proboscis, proboscis armature and receptacle, lemnisci, and reproductive organs. A detailed study of specimens from South Africa and descriptions reported from elsewhere in Africa revealed marked differences that clearly distinguish the African material as new species. The African specimens are pseudo-segmented and flattened, the proboscis has two prominent apical pores, sensory pits are prevalent throughout the trunk, the posterior end of the female is broad with dorso-terminal dome-like extension opposite the subterminal gonopore, and the eggs are large. The Asian specimens from Indonesia and elsewhere are cylindrical and non-segmented, the proboscis lacks prominent apical pores, sensory pits are rare on the trunk, the posterior end of the female is pointed with a terminal gonopore, and the eggs are markedly smaller. We used DNA sequence from one mitochondrial gene (cytochrome oxidase subunit I) and one nuclear gene (18S ribosomal RNA) to infer the phylogenetic relationships of M. africanus and M. gallinarum and selected Acanthocephala. Medioryhnchus is monophyletic and M. africanus and M. gallinarum are allopatric sister species (9.7% sequence divergence). All findings indicate that M. africanus should be ranked as a separate species.

The morphology and histopathology of Nephridiacanthus major (Acanthocephala: Oligacanthorhynchidae) from hedgehogs in Iran.

The morphology of Nephridiacanthus major (Bremser 1811 in Westrumb 1821) Golvan, 1962 collected from the long-eared hedgehog Hemiechinus auritus (Gmelin 1770) and the Eastern European hedgehog Erinaceus concolor Martin, 1838 (Erinaceidae) is described using SEM for the first time. This acanthocephalan was previously described from hedgehogs in Europe, Asia, and Africa. Measurements of specimens from Iran, Bulgaria, Germany, Central Asia, Morocco, and Egypt show considerable variations in the size of the trunk, proboscis, proboscis hooks and receptacle, and eggs. The SEM studies add new perspectives to its morphology. Features observed for the first time include the near terminal position and shape of the female gonopore and orifice, among others. Histopathological studies for this species are reported for the first time. Tissue sections show extensive damage near the proboscis with hemorrhaging and formation of collagenous connective tissue, compression of the intestinal mucosa, obstruction of intestinal lumen, and extensive necrosis of host epithelial tissue.