Hypoglycemia associated with disseminated metastatic tonsillar squamous cell carcinoma producing insulin‐like growth factor‐I in a Pomeranian dog

A 13‐year‐old spayed female Pomeranian dog was presented for persistent, severe hypoglycemia (37 mg/dL; reference interval [RI] 75‐128 mg/dL). Progressive nonregenerative anemia (hematocrit 23.3%‐15.9%; RI 37.0%‐55.0%) and severe thrombocytopenia (36 000/µL; RI 200‐500 000/µL) were also noted. The serum insulin concentration was low (0.24 ng/mL; RI 0.302‐1.277 ng/mL). Computed tomography revealed multiple splenic nodules (1‐6 mm in diameter) and several hepatic nodules (7.6, 12 mm in diameter). Ultrasound‐guided fine‐needle aspiration of the splenic and hepatic nodules revealed low numbers of epithelial cells with mild cellular atypia, suggestive of a metastatic epithelial tumor, but the primary site was unknown at that time. On careful oral examination under general anesthesia, an enlarged right tonsil was noted grossly, and histopathologic examination of the tonsil diagnosed squamous cell carcinoma. Bone marrow aspirates and biopsies of the splenic and hepatic nodules were performed; all samples were diagnosed as metastatic carcinoma on histopathologic examination. No nodules were present in the pancreas, despite careful palpation during exploratory laparotomy. On immunohistochemistry, the neoplastic cells were positive for cytokeratin AE1/3 and insulin‐like growth factor (IGF)‐I but were negative for chromogranin A, PGP9.5, insulin, and inconclusive for IGF‐II. This is the first report of a primary IGF‐I‐producing squamous cell carcinoma in the tonsil of a dog with metastases to bone marrow, liver, and spleen, resulting in hypoglycemia.     

Microbial biomass and nitrogen transformations in surface soils strongly acidified by volcanic hydrogen sulfide deposition in Osorezan, Japan

Volcanic acidification has created unique ecosystems that have had to adapt to the acidic environments in volcanic regions. To characterize the primary microbial properties of strongly acidified soils in such environments, we investigated microbial biomass, nitrogen transformations and other relevant chemical properties in the surface soils of solfatara and forests from Osorezan, a typical volcanic region in Japan, and compared the results to common Japanese forest soils. Soil microbial biomass C (MBC) and N (MBN) were determined using the chloroform fumigation–extraction method. Potential net N mineralization and net nitrification were measured in aerobic laboratory incubations. Long-term acidification in the Osorezan soils by volcanic hydrogen sulfide deposition caused low soil pH (3.0–3.8), base cation deficiency and increased concentrations of toxic ions such as Al³⁺. The proportions of MBC to total carbon (MBC/TC ratio) and MBN to total nitrogen (MBN/TN ratio) were lower than those in common Japanese forest soils. The extreme acidic conditions may have inhibited microbial survival in the Osorezan acid soils. Net N mineralization occurred at rates comparable to those in common Cryptomeria japonica forest soils, probably because of the presence of acid-tolerant soil microorganisms. Net nitrification was completely inhibited and autotrophic ammonia oxidizers were not detected by the MPN method. The inhibition of nitrification prevents nitrogen leaching from the soils, thus maintaining a nitrogen cycle in the volcanic acid region in which     (and NH₃) is recycled among microorganisms and plants.     

Transepithelial transport of chlorogenic acid, caffeic acid, and their colonic metabolites in intestinal caco-2 cell monolayers

Both chlorogenic and caffeic acids exhibited nonsaturable transport in Caco-2 cells, whereas caffeic acid also showed proton-coupled polarized absorption. Thus, the absorption efficiency of caffeic acid was greater than that of chlorogenic acid. Polarized transport of caffeic acid was inhibited by substrates of MCT such as benzoic and acetic acids. Almost all of the apically loaded chlorogenic and caffeic acid was retained on the apical side, and the transepithelial flux was inversely correlated with the paracellular permeability of Caco-2 cells. These results indicate that transport was mainly via paracellular diffusion, although caffeic acid was absorbed to a lesser extent by the monocarboxylic acid transporter (MCT). Furthermore, m-coumaric acid and 3-(m-hydroxyphenyl)propionic acid, the main metabolites of chlorogenic and caffeic acid by colonic microflora, competitively inhibited the transport of fluorescein, a known substrate of MCT. This suggests that their absorption could also be mediated by MCT. These findings have exemplified the physiological importance of MCT-mediated absorption in both phenolic acids per se and their colonic metabolites.     

Tea polyphenols inhibit the transport of dietary phenolic acids mediated by the monocarboxylic acid transporter (MCT) in intestinal Caco-2 cell monolayers

It was previously reported that a fluorescent marker dye, fluorescein, is transported via the monocarboxylic acid transporter (MCT). Fluorescein transport was competitively inhibited by MCT substrates such as ferulic and salicylic acids. Tea polyphenols, in particular, epigallocatechin gallate (EGCg) and epicatechin gallate (ECg), inhibited the transport of fluorescein. Tea polyphenols also inhibited the transport of salicylic and ferulic acids, suggesting tea polyphenols might be substrates of MCT. However, the transepithelial flux of tea polyphenols was much lower than that of the MCT substrates and was inversely correlated with the paracellular permeability of Caco-2 cell monolayers. These findings suggest that tea polyphenols are not substrates but inhibitors of MCT. Furthermore, the transepithelial transport of these polyphenols is mainly via paracellular diffusion. However, directional transport of ECg and EGCg from the basolateral to the apical side was observed, indicating that the behavior of tea polyphenols in the intestinal epithelium is complex.     

Changes in Chemical Species of Copper Added to Brown Forest Soil in Japan

Heavy metals are present as various chemical species in soil environments. Chemical species of heavy metals determinate their behavior in soil, so they must be more important concern than total contents in treating problems about heavy metal polluted soil. Microbial activity is thought to be one of the factors that affect chemical speciation of heavy metals. Although some possible microbial mechanisms have been suggested, their effects on heavy metals have not previously been thoroughly studied. In this study, we investigated whether microorganisms contribute to the fractionation of Cu in a forest soil. Copper-enriched, sterilized forest soil samples were buried at their places of origin, and subsamples were collected for analysis 3, 10, 17, 30, 60, and 90 days after burial. Tessier’s sequential extraction was employed to analyse Cu fractionation. Until two months after burial, the added Cu remained nearly unchanged. But a dramatic change was found in soil subsamples after three months. In both the A and B horizons, the carbonate fraction of Cu increased and the residual fraction decreased. We also found that clay minerals and persistent organic matter are important binding substances of Cu in the residual fraction. The change from residual to carbonate fraction could be caused by microbial activity.     

Assessment of potentially reactive pools of aluminum in Andisols using a five-step sequential extraction procedure

A five-step sequential extraction procedure was developed for the partitioning of soil aluminum (Al) into five fractions: exchangeable, weakly organic bound, strongly organic bound, inorganic non-crystalline and crystalline. The results obtained by the sequential extraction method for Al were compared with those estimated from single extractions using identical operating conditions applied in each individual sequential fraction. The Al content in the extracts was measured by inductively coupled plasma optical emission spectrometry. The results indicated that the first four steps [potassium chloride (KCl), copper chloride (CuCl₂), sodium pyrophosphate (Na₄P₂O₇), acid ammonium oxalate ((NH₄)₂C₂O₄)] in the sequential procedure could be as effective as single extraction methods at estimating exchangeable Al, weakly organic bound Al, strongly organic bound Al, and non-crystalline Al. However, the crystalline Al content by sequential procedure was not in agreement with single extraction procedures. Further, the sequential extractions resulted in more consistent estimates of the aluminum/silicon (Al/Si) molar ratio and allophane and crystalline Al contents than single extractions method. Results of X-ray diffraction on the soil samples confirm the presence of appreciable amounts of at least three types of crystalline minerals, including gibbsite, vermiculite or halloysite. Our result implies that the sequential method can be more reliable in estimating the various Al forms in Andisols.     

Dietary vitamin A requirements of juvenile Japanese flounder Paralichthys olivaceus

A 70‐day feeding experiment was conducted to assess the dietary vitamin A (VA) requirements of juvenile Japanese flounder (Paralichthys olivaceus). Six semi‐purified diets with VA supplementations of 0, 5000, 10 000, 15 000, 20 000 and 25 000 IU kg^?1 were fed twice a day to triplicate groups of 20 juveniles per tank with an initial weight of 1.59 ± 0.01 g (mean ± SE). Weight gain (WG) and specific growth rate (SGR) increased as dietary VA increased up to 10 000 IU kg^?1. Significantly lower WG and SGR were observed for the 0 IU kg^?1 treatment than for treatments of 5000, 10 000 and 15 000 IU kg^?1. Highest WG and SGR were observed in fish fed 10 000 IU kg^?1; slightly lower values were recorded in fish fed 15 000, 20 000 or 25 000 IU kg¹. No significant difference was observed in survival rate among treatments. Whole body total lipid was significantly higher in fish fed 0 and 5000 IU kg^?1 than for other levels. Reduced growth and small livers were observed as signs of VA deficiency in fish fed 0 IU kg^?1. Slightly reduced growth and pale fragile livers were observed as effects of VA excess in fish fed 25 000 IU kg^?1. Total retinol contents in liver and eyes increased with increasing levels of dietary VA. No retinol was detected in livers, and significantly lower total retinol content was observed in eyes, of fish fed 0 IU kg^?1. WG analysed by the broken line method indicated that an optimum dietary VA requirement of 9000 IU kg^?1.