Renal Infiltration of Macrophages in Canine and Feline Chronic Kidney Disease.

During the progression of chronic kidney disease (CKD), macrophage infiltration is a crucial event leading to tubulointerstitial fibrosis. In the present study, macrophages infiltrating renal tissue in dogs and cats with CKD were analysed immunohistochemically. Iba-1 was used as a pan-macrophage marker, CD204 was used as a marker of M2 macrophages and tumour necrosis factor (TNF)-α was used as a marker of M1 macrophages. Signals for Iba1 and CD204 were observed in the interstitium of all tested kidney samples. In dogs, the signals were diffusely scattered. In cats, both diffuse and focal signals were observed. Cells that were positive for Iba1 and CD204 were also observed in the tubular lumina in cats. Co-expression of Iba1 and CD204 was also observed in the infiltrating cells by immunofluorescence labelling, and these cells were negative for TNF-α. By quantitative analysis, the indices for Iba1- and CD204-positive cells were significantly correlated with the concentrations of plasma creatinine and/or urea and the extent of interstitial fibrosis in both dogs and cats. These results demonstrated that renal infiltration of M2 macrophages plays an important role in the progression of CKD in dogs and cats. The distribution pattern of the kidney-infiltrating macrophages was unique in cats and may be associated with a cat-specific renal fibrotic process.

Biotransport of metallic trace elements from marine to terrestrial ecosystems by seabirds.

Physical systems, such as currents and winds, have traditionally been considered responsible for transporting contaminants. Although evidence is mounting that animals play a role in this process through their movements, we still know little about how such contaminant biotransport occurs and the extent of effects at deposition sites. In the present study, we address this question by studying how rhinoceros auklets (Cerorhinca monocerata), a seabird that occurs in immense colonies (∼300 000 pairs at our study site, Teuri Island), affect contaminant levels at their colony and at nearby sites. More specifically, we hypothesize that contaminants are transported and deposited by seabirds at their colony and that these contaminants are passed on locally to the terrestrial ecosystem. To test this hypothesis, we analyzed the concentration of 9 heavy metal and metalloids, as well as δ13 C and δ15 N stable isotopes, in bird tissues, plants, and soil, both within and outside of the colony. The results show that rhinoceros auklets transport marine-derived mercury (Hg), possibly from their wintering location, and deposit Hg via their feces at their breeding site, thereby contaminating plants and soils within the breeding colony. The present study confirms not only that animals can transport contaminants from marine to terrestrial ecosystems, potentially over unexpectedly long distances, but also that bird tissues contribute locally to plant contamination. Environ Toxicol Chem 2019;38:106-114. © 2018 SETAC.

Changes in Renal Peritubular Capillaries in Canine and Feline Chronic Kidney Disease.

Renal capillary rarefaction is a crucial event that leads to tubulointerstitial damage during the progression of chronic kidney disease (CKD). In the present study, changes in CD34-positive renal capillaries were investigated in dogs and cats with CKD. A significant decrease in CD34-positive capillaries was observed in canine diseased kidneys, even at the early stage of disease. In cats, CD34-positive capillaries were well preserved in the diseased kidneys, with no link to the severity of CKD. Renal capillary rarefaction might be a trigger event that leads to the progression of CKD in dogs, rather than in cats.

Human Health Risk from Consumption of Marine Fish Contaminated with DDT and Its Metabolites in Maputo Bay, Mozambique.

Many countries with incidence of malaria, including those surrounding Maputo Bay, use dichloro-diphenyl-trichloroethane (DDT) to reduce mosquitoes. This study is the first to estimate the human health risk associated with consumption of marine fish from Maputo Bay contaminated with DDTs. The median for ∑DDTs was 3.8 ng/g ww (maximum 280.9 ng/g ww). The overall hazard ratio for samples was 1.5 at the 75th percentile concentration and 28.2 at the 95th percentile. These calculations show increased potential cancer risks due to contamination by DDTs, data which will help policy makers perform a risk-benefit analysis of DDT use in malaria control programs in the region.

A glycomics approach to discover novel renal biomarkers in birds by administration of cisplatin and diclofenac to chickens.

Avian species have a unique renal structure and abundant blood flow into the kidneys. Although many birds die due to nephrotoxicity caused by chemicals, there are no early biomarkers for renal lesions. Uric acid level in blood, which is generally used as a renal biomarker, is altered when the kidney function is damaged by over 70%. Therefore, early biomarkers for kidney injury in birds are needed. In humans, glycomics has been at the forefront of biological and medical sciences, and glycans are used as biomarkers of diseases, such as carcinoma. In this study, a glycomics approach was used to screen for renal biomarkers in chicken. First, a chicken model of kidney damage was generated by injection of diclofenac or cisplatin, which cause acute interstitial nephritis (AIN) and acute tubular necrosis (ATN), respectively. The nephrotoxicity levels were determined by a blood chemical test and histopathological analysis. The plasma N-glycans were then analyzed to discover renal biomarkers in birds. Levels of 14 glycans increased between pre- and post administration in kidney-damaged chickens in the diclofenac group, and some of these glycans had the same presumptive composition as those in human renal carcinoma patients. Glycan levels did not change remarkably in the cisplatin group. It is possible that there are changes in glycan expression due to AIN, but they do not reflect ATN. Although further research is needed in other species of birds, glycans are potentially useful biomarkers for AIN in avian species.

Concentrations and human health risk assessment of DDT and its metabolites in free-range and commercial chicken products from KwaZulu-Natal, South Africa.

Organochlorine pesticides such as dichlorodiphenyltrichloroethane (DDT) have been used in agriculture and for disease control purposes over many decades. Reports suggest that DDT exposure may result in a number of adverse effects in humans. In the KwaZulu-Natal Province of South Africa, DDT is sprayed annually in homes (indoor residual spraying) to control the mosquito vector of malaria. In the northern part of the Province, samples of free-range chicken meat (n = 48) and eggs (n = 13), and commercially produced chicken meat (n = 6) and eggs (n = 11), were collected and analysed. Of the free-range chicken meat samples, 94% (45/48) contained DDTs (ΣDDTs median 6.1 ng/g wet weight [ww], maximum 79.1 ng/g ww). Chicken egg contents were also contaminated (ΣDDTs in free-range eggs median 9544 ng/g ww, maximum 96.666 ng/g ww; and in commercial eggs median 1.3 ng/g ww, maximum 4.6 ng/g ww). The predominant DDT congener detected was p,p'-DDE in both free-range meat (>63%) and eggs (>66%), followed by p,p'-DDT and then p,p'-DDD. Based on estimated daily intake values, calculated human risk ratio (carcinogenic) values were >1 for DDTs detected in both free-range chicken products. Consumption of free-range eggs poses a particularly high health risk.

Characterization of equine cytochrome P450: role of CYP3A in the metabolism of diazepam.

Research on drug metabolism and pharmacokinetics in large animal species including the horse is scarce because of the challenges in conducting in vivo studies. The metabolic reactions catalyzed by cytochrome P450s (CYPs) are central to drug pharmacokinetics. This study elucidated the characteristics of equine CYPs using diazepam (DZP) as a model compound as this drug is widely used as an anesthetic and sedative in horses, and is principally metabolized by CYPs. Diazepam metabolic activities were measured in vitro using horse and rat liver microsomes to clarify the species differences in enzyme kinetic parameters of each metabolite (temazepam [TMZ], nordiazepam [NDZ], p-hydroxydiazepam [p-OH-DZP], and oxazepam [OXZ]). In both species microsomes, TMZ was the major metabolite, but the formation rate of p-OH-DZP was significantly less in the horse. Inhibition assays with a CYP-specific inhibitors and antibody suggested that CYP3A was the main enzyme responsible for DZP metabolism in horse. Four recombinant equine CYP3A isoforms expressed in Cos-7 cells showed that CYP3A96, CYP3A94, and CYP3A89 were important for TMZ formation, whereas CYP3A97 exhibited more limited activity. Phylogenetic analysis suggested diversification of CYP3As in each mammalian order. Further study is needed to elucidate functional characteristics of each equine CYP3A isoform for effective use of diazepam in horses.

Partial cloning of CYP2C23a genes and hepatic protein expression in eight representative avian species.

Large interspecies differences in avian xenobiotic metabolism have been revealed by microsome-based studies, but specific enzyme isoforms in different bird species have not yet been compared. We have previously shown that CYP2C23 genes are the most induced CYP isoforms in chicken liver. In this study, we collected partial CYP2C23a gene sequences from eight avian species (ostrich, blue-eared pheasant, snowy owl, great-horned owl, Chilean flamingo, peregrin falcon, Humboldt penguin, and black-crowned night heron) selected to cover the whole avian lineage: Paleognathae, Galloanserae, and Neoaves. Genetic analysis showed that CYP2C23 genes of Galloanserae species (chicken and blue-eared pheasant) had unique characteristics. We found some duplicated genes (CYP2C23a and CYP2C23b) and two missing amino acid residues in Galloanserae compared to the other two lineages. The genes have lower homology than in other avian lineages, which suggests Galloanserae-specific rapid evolutionary changes. These genetic features suggested that the Galloanserae are not the most representative avian species, considering that the Neoaves comprise more than 95% of birds. Moreover, we succeeded in synthesizing an antipeptide polyclonal antibody against the region of CYP2C23 protein conserved in avians. However, comparative quantitation of CYP2C23 proteins in livers from six species showed that expression levels of these proteins differed no more than fourfold. Further study is needed to clarify the function of avian CYP2C23 proteins.

The effects of ginseng radix rubra on human vascular endothelial cells.

The effect of Ginseng Radix Rubra (Red ginseng) on human vascular endothelial cells was examined. Red ginseng was found to promote the proliferation of vascular endothelial cells, inhibit the production but promote the decomposition of endothelin, which is known to constrict blood vessels and raise blood pressure as well as accelerated the synthesis of nitric oxide, which is known to have an angio-tonic effect. Furthermore, Red ginseng was observed to increase the production of Interleukin 1 beta, which is known to play important roles in the homeostatic activities of the human body such as immunity and inflammation as well as increasing the production of tissue plasminogen activators, which suppress the formation of thrombin in the blood coagulation and fibrinolysis mechanisms. It is suggested that Red ginseng has the effect of accelerating endothelial cells proliferation and of promoting physiological activities.

Inhibition by singlet molecular oxygen of the vascular reactivity in rabbit mesenteric artery.

1. The effects of reactive oxygen intermediates derived from photoactivated rose bengal on the vascular reactivity have been evaluated in rabbit mesenteric artery ring preparations. The artery rings were exposed to xanthene dye rose bengal (50 nM) illuminated (6,000 lux) at 560 nm for 30 min. Spin trapping studies with 2,2,6,6-tetramethylpiperidine (TEMP) and 5,5-dimethyl-1-pyrroline-N-oxide (DMPO) with electron spin resonance spectrometry were also conducted in solution (and not within tissues) to determine quantitatively the reactive oxygen species generated from photoactivated rose bengal. 2. Contraction of the ring preparations induced by noradrenaline (10(-8) to 10(-4) M) was attenuated by previous exposure to photolysed rose bengal; the observation that the pD2 decreased without a significant reduction in maximum tension generation is consistent with the view that receptor dysfunction may be involved in the effect of photolysed rose bengal. 3. Prior exposure to photolysed rose bengal of the ring preparations inhibited the endothelium-dependent relaxation evoked by acetylcholine (10(-6) M) and calcium ionophore A23187 (10(-7) M), but not the endothelium-independent relaxation evoked by nitroglycerin (10(-6) M). 4. A variety of scavengers, superoxide dismutase (33 units ml-1), catalase (32 units ml-1) and 1,3-dimethyl-2-thiourea (DMTU, 10 mM), which should eliminate the superoxide anion radical, H2O2 and the hydroxyl radical, had no effect on the attenuated responses to noradrenaline and acetylcholine induced by photolysed rose bengal. In contrast, the inhibition of the observed effect of photolysed rose bengal was obtained with addition of histidine (25 mM), a singlet molecular oxygen quencher. 5. It was found that photolysis of rose bengal from a 1:2:2:1 quartet, characteristic of the hydroxyl radical-DMPO spin adduct, which was effectively blunted by DMTU, superoxide dismutase and catalase whereas histidine was ineffective. The results of the electron spin resonance study also showed that a singlet molecular oxygen was produced by photoactivation of rose bengal; this was detected as singlet oxygen-TEMP product (TEMPO; 2,2,6,6-tetramethylpiperidine-N-oxyl). The formation of the TEMPO signal was strongly inhibited by histidine, but not by DMTU, superoxide dismutase and catalase. 6. It is suggested that the superoxide anion radical, H2O2 and hydroxyl radical are formed in addition to singlet molecular oxygen, and the data obtained from the present study indicate that singlet molecular oxygen is one of the most destructive oxygen species. Endothelium-dependent relaxation is quite vulnerable to singlet molecular oxygen. Singlet oxygen also depresses noradrenaline-induced contraction possibly via alpha-adrenoceptor dysfunction. This, in turn, may lead to vascular incompetence.

The effects of ninjinyoeito on cultured venous endothelial cells.

The effects of Ninjinyoeito on endothelial cells in human umbilical veins was examined. We found that Ninjinyoeito accelerates the inhibition and decomposition of Endothelin-1 production, which is known to constrict veins and raise blood pressure, and promote the synthesis of nitric oxicide which is known to have a vasophypotonic effect. We also found that Ninjinyoeito accelerates Interleukin 1-beta (IL-1 beta) production. IL-1 beta is a substance that plays important roles in maintaining the homeostasis of living organisms in immunity and inflammation, for example, as well as in the production of tissue plasminogen activators, which are known to have an anti-thrombotic effect on blood coagulation and on the fibrinolysis mechanism. Thus, it is suggested that Ninjinyoeito is effective in accelerating the physiological functions of human vein endothelial cells in vitro.

[Peripheral leukocyte adhesion molecules in patients of Behçet's disease associated with active ocular lesions].

The sequential cascades in leukocyte adhesion and migration are important events in the development of inflammatory and immune responses in Behçet's disease. In an attempt to clarify the relation of active ocular lesions to inflammatory reactions covering entire body, we have detected LECAM-1, Mac-1 and CD44 expressed on the peripheral leukocytes in 24 Behçet patients and 15 healthy adults by flow cytometry. LECAM-1 and CD44 expression was dramatically decreased from the polymorphonuclear leukocytes (PMN) upon ocular attacks, whereas the changes in Mac-1 appeared not so striking. This tendency had continued into the recovery stage of ocular inflammation. T lymphocytes showed, on the other hand, no considerable variability in regard to the expression of cell surface LECAM-1, CD44 and also of Mac-1 even at the peak of active ocular inflammation. The results suggest that interaction between PMN and primed vascular endothelial cells might be apt to precede functional modifications of T lymphocytes, and it is also supposed that facilitation of activated PMN recruitment plays an essential role on ocular inflammation in Behçet's disease.

The effect of sho-saiko-to on concentration of vitamin E in serum and on granuloma formation in carrageenin cotton pellet-induced granuloma rats.

The effect of Sho-saiko-to on the concentration of vitamin E in serum and on the granuloma formation in Carrageenin cotton pellet-induced rats was investigated. As a result, in the granuloma rats of Sho-saiko-to group, a significantly improved inhibitory effect on granuloma formation and a higher concentration of vitamin E in serum, cholesterol and phospholipid were observed compared to the control group. Despite this lipid-increasing action by Sho-saiko-to, the concentration of serum lipid peroxide was significantly lower than in the control group. Furthermore, a significant negative correlation between the concentration of vitamin E and granuloma weight was observed. These results suggest that vitamin E plays an important role in promoting the anti-inflammatory effect of Sho-saiko-to.

The effect of ninjinyoeito on human aorta endothelial cells.

We studied the effect of Ninjinyoeito in relation to the endothelial cells of the human aorta. Ninjinyoeito produced a cell proliferation acceleration effect, resulting in suppression of the synthesis and acceleration of the disintegration of endothelin, which acts as a vasoconstrictor and vasopressor. Also, acceleration of the synthesis of prostacyclin, which shows antithrombosis and platelet coagulation suppressing effects, was observed. The results suggest that Ninjinyoeito accelerates physiological function of in vitro human aorta endothelial cells.

Is personality involved in the expression of dysmenorrhea in patients with endometriosis?

OBJECTIVE: To investigate whether a relationship exists between personality and expression of pain in women with endometriosis. STUDY DESIGN: With use of the Rosenzweig picture frustration study, the personality of women with endometriosis was assessed. These results were then correlated with the manifestation of pain. RESULTS: Women without dysmenorrhea tended to be less assertive compared with women who complained of dysmenorrhea and women without endometriosis. Those women without dysmenorrhea showed a destructive attitude in face of a problem. CONCLUSIONS: Personality affected the expression of pain. Pain caused by endometriosis was influenced by each personality type.

The effects of ninjinyoeito on human vascular endothelial cells.

The effects of Ninjinyoeito on endothelial cells from a human umbilical vein were examined. As a result, the maximum cell density obtained after the addition of Ninjinyoeito was found to be 400 ng/ml, and its inhibitory effect on the development of endothelial injury was also observed. In addition, the levels of LDH and Al-p, which are parameters decreasing with aging, were significantly high. Judging from these results, Ninjinyoeito presumably exerts an anti-aging effect by inhibiting the metabolic aging of cells.

The effect of orengedokuto on experimentally-inflamed rats.

The inhibitory effect of Orengedokuto on lipid peroxide and sialidase was investigated using rats affected with carrageenin cotton pellet-induced granuloma and adjuvant arthritis. As a result, in the case of rats with carrageenin cotton pellet-induced granuloma, a significant inhibitory effect on granuloma formation was observed in the Orengedokuto treated rats (150 mg/kg/day) which showed a decrease in serum lipid peroxide (p < 0.001) and an increase in acid soluble glycoprotein (p < 0.001). In the case of adjuvant-induced arthritic rats, however, the condition of the arthritis was not improved at all, and was even aggravated in spite of the decrease in serum lipid peroxide.

The effect of sho-saiko-to on the concentration of acid soluble glycoprotein in serum and on granuloma formation in carrageenin cotton pellet-induced granuloma rats.

The effect of sho-saiko-to on the concentration of acid soluble glycoprotein in serum and on the granuloma formation in carrageenin cotton pellet-induced rats was investigated. As a result, a significant negative correlation between the concentration of acid soluble glycoprotein and granuloma weight was observed. Furthermore, both the concentration of acid soluble glycoprotein and the inhibition rate of the granuloma formation were significantly higher in the sho-saiko-to group than in the control group. These results suggest that acid soluble glycoprotein plays an important role in promoting the anti-inflammatory effect of sho-saiko-to.

[Clinical significance of serum dipeptidyl aminopeptidase IV in rats with adjuvant arthritis].

Serum dipeptidyl aminopeptidase IV (DAP IV) activity in rats with adjuvant arthritis (AA) were studied. DAP IV activity was significantly low in AA rats (37.7 +/- 14.1 mU/ml, n = 15) compared with control (63.7 +/- 9.0 mU/ml, n = 8) and indomethacin-treated (IDM) AA rats (55.6 +/- 14.2 mU/ml, n = 15). In addition, serum DAP IV activity was inversely correlated with hindpaw arthritis score both in AA rats and IDM-AA rats. Among serum inflammatory index examined, acid soluble glycoprotein, sialic acid were inversely correlated and albumin was positively correlated with DAP IV activity in AA and IDM-AA rats. The role of serum DAP IV remains unknown, but, it seems possible that DAP IV might modify the clinical course of AA rats in connection with its effects on the immune regulatory system.