A review of anaemia of inflammatory disease in dogs and cats.

Anaemia of inflammatory disease is a common cause of anaemia in routine veterinary practice. It is most often mild to moderate, normocytic, normochromic and non-regenerative. Shortened red cell life span, inhibition of iron metabolism and impaired bone marrow response to erythropoietin all contribute to its development. Although anaemia of inflammatory disease is a well-known cause of anaemia in dogs and cats, there is a lack of epidemiological information because specific diagnostic criteria have not been established in veterinary species. Anaemia of inflammatory disease is associated with a poor outcome in various disease states in human medicine; however, its clinical significance and treatment in veterinary medicine are not well understood. This review article describes anaemia of inflammatory disease in dogs and cats and considers its potential significance.

Fish collagen is an important panallergen in the Japanese population.

Collagen was identified as a fish allergen in early 2000s. Although its allergenic potential has been suggested to be low, risks associated with collagen as a fish allergen have not been evaluated to a greater extent. In this study, we aimed to clarify the importance of collagen as a fish allergen. Our results showed that 50% of Japanese patients with fish allergy had immunoglobulin E (IgE) against mackerel collagen, whereas 44% had IgE against mackerel parvalbumin. IgE inhibition assay revealed high cross-reactivity of mackerel collagen to 22 fish species (inhibition rates: 87-98%). Furthermore, a recently developed allergy test demonstrated that collagen triggered IgE cross-linking on mast cells. These data indicate that fish collagen is an important and very common panallergen in fish consumed in Japan. The high rate of individuals' collagen allergy may be attributable to the traditional Japanese custom of raw fish consumption.

Prevalence of intestinal parasites and genotyping of Giardia intestinalis in pet shop puppies in east Japan.

The current study examined the prevalence of intestinal parasites and genotypes of Giardia intestinalis in puppies from nine pet shops in east Japan. Fresh fecal samples from 1794 puppies (≦3 months old) were collected on one occasion. Giardia spp. was examined for specific coproantigen using ELISA kit (SNAP®Giardia, IDEXX Laboratories, Inc., USA). Other intestinal parasites were detected microscopically using the formalin-ethyl acetate sedimentation technique. Genotyping was determined for the random 29 stool samples identified as Giardia spp. positive using PCR and direct sequencing of the glutamate dehydrogenase (gdh) gene. Overall prevalence of protozoan Giardia spp. and Cystoisospora spp. revealed 23.4% and 11.3%, respectively. Prevalence of ascarids, Strongyloides spp. and hookworms were recorded 1.8%, 1.1% and 0.1%, respectively. Protozoan Giardia spp. and Cystoisospora spp., thus, represent important pathogens among pet shop puppies. All genotyped G. intestinalis isolates were belonged to assemblage C or D, identified as dog-specific genotypes. Zoonotic assemblage A and B were not demonstrated. The result suggests that the risk of zoonotic transmission of G. intestinalis from pet shops puppies to humans may be quite low in Japan.

N-Acetyltransferase2 genotype correlated with isoniazid acetylation in Japanese tuberculous patients.

Isoniazid (INH) is metabolized by polymorphic N-acetyltransferase2 (NAT2). In the present study, the relationship between the NAT2 genotype and the INH acetylator phenotype was examined in Japanese tuberculous patients and compared with healthy subjects. Subjects were classified according to the genotyping into NAT2*5B (allele4), NAT2*6A (allele3) and NAT2*7B (allele2), using the PCR-RFLP method. Twelve healthy subjects and 7 tuberculous patients participated in the INH acetylator phenotyping study, in which each subject was administered an oral dose of INH, followed by urine sampling for 24 h. Urinary concentrations of INH and N-acetylisoniazid (AcINH) were measured by the HPLC method. The urinary recoveries of INH (% of dose) in healthy subjects in relation to NAT2 genotyping were as follows: 6.4+/-2.2 in the homozygotes for the wild-type allele, 10.7+/-2.2 in the compound heterozygotes for the mutant allele, and 38.6+/-6.4 in the homozygotes for the mutant allele. In the patients study, the findings in the corresponding three groups were 4.0+/-1.7, 8.8 and 18.3+/-9.3. Although no significant difference was found because of the lower systemic exposure of INH in patients compared with healthy subjects, there were differences in the disposition kinetics of INH between subjects with and without mutations in the NAT2 gene, and these findings were observed not only in healthy subjects but also in patients who had comedicated drugs and hepatic dysfunctions. The findings indicated that the metabolism of INH by NAT2 is clearly impaired in subjects with mutations in the NAT2 gene, and thus genotyping for three NAT2 point mutations was adequate to predict the metabolism of INH in Japanese tuberculous patients as well as healthy subjects. This NAT2 genotyping could become a useful alternative to TDM for INH.

Genotyping of N-acetylation polymorphism and correlation with procainamide metabolism.

We studied the genotypes of polymorphic N-acetyltransferase (NAT2) in 145 Japanese subjects by the polymerase chain reaction-restriction fragment length polymorphism method. The rapid-type NAT2*4 was expressed at a higher frequency (68.6%) than the slow-type genes with specific point mutations (NAT2*6A, 19.3%; NAT2*7B, 9.7%; NAT2*5B, 2.4%). The frequency of NAT2* genotypes consisted of 44% of a homozygote of NAT2*4, 49% of a heterozygote of NAT2*4 and mutant genes, and 7% of a combination of mutant genes. The metabolic activity for procainamide to N-acetylprocainamide was measured in 11 healthy subjects whose genotype had been determined. Although the acetylation activity substantially varied interindividually, the variability was considerably reduced after classification according to the genotype. The N-acetylprocainamide/procainamide ratio in urinary excretion was 0.60 +/- 0.17 (mean +/- SD) for those with NAT2*4/*4, 0.37 +/- 0.06 for NAT2*4/*6A, 0.40 +/- 0.03 for NAT2*4/*7B, and 0.17 for NAT2*6A/*7B. The results indicated that the NAT2* genotype correlates with acetylation of procainamide.

Relationship between serum concentration and daily dose of dantrolene in cerebral palsy patients.

Serum concentration of dantrolene and its active metabolite, 5-hydroxydantrolene were determined in 27 cerebral palsy patients. Correlation coefficients between the oral dose of dantrolene and serum levels of dantrolene and its major metabolite, 5-hydroxydantrolene were rather small in cerebral palsy patients. The mean half-times of dantrolene and 5-hydroxydantrolene were 3.41 (n = 6) and 4.00 (n = 5) hours, respectively. These values were about a half of those reported earlier [Lietman et al., 1974; Meyler et al., 1979; Flewellen et al., 1983]. The poor correlation between serum level and dose may be due to the variation in an extent of oral dantrolene availability.

Regulation of receptors and digestive activity toward synthesized formyl-chemotactic peptide in human polymorphonuclear leukocytes.

A receptor binding and digestive activity of human polymorphonuclear leukocytes (PMNs) toward formyl-methionyl-leucyl-[3H]phenylalanine (3H-FMLP) was examined with the following results: Up- and down-regulation and recovery of 3H-FMLP binding activity were demonstrated. Both intact PMN and a lysate prepared from them cleaved the carboxyl terminal amino acid (phenylalanine) of 3H-FMLP. The digestive activity decreased as the receptor binding was inhibited by n-ethylmaleimide and 4-chloromercuribenzoate. Little digestive activity was found in the supernatant from PMN stimulated by FMLP. The released phenylalanine was found in the pellet and supernatant of PMNs. Digestive activity with cathepsin A-like characteristics was found in the lysate of PMN. These observations suggest that FMLP is internalized in lysosomes in a receptor-mediated manner and cleaved by the cathepsin A-like enzyme, the free phenylalanine is released extracellularly, and a part of the dissociated receptors with FMLP may return to the surface or to an intracellular receptor pool. Another finding was that the digestive activity of the lysate of cord blood granulocytes was decreased compared with that of adult blood granulocytes. This decrease may explain in part the impaired chemotaxis of cord blood granulocytes.

Hyperimmunoglobulin-E-associated recurrent infection syndrome accompanied by chemotactic inhibition of polymorphonuclear leukocytes and monocytes.

An 8-yr-old girl with a history of severe recurrent infections including perinephritic, pulmonary, and hepatic abscesses had elevated serum IgE levels. Her serum inhibited chemotaxis of polymorphonuclear leukocytes (PMN) and monocytes. Exchange blood transfusion or plasma exchange at the time of severe infection resulted in normalization of chemotactic activity of PMN shown by the skin window method. Although this effect became negative 1 wk after the treatment, the procedures improved her clinical course. The patient's serum, obtained by exchange blood transfusion, 1) inhibited normal PMN chemotaxis toward cultured supernatant of E. coli, zymosan-activated serum, and formyl methionyl-leucyl-phenylalanine (f . Met-Leu-Phe), a synthetic chemotactic peptide; 2) inhibited monocyte chemotaxis, 3) showed an absence of digestive activity of f . Met-Leu-Phe, 4) was heat stable at 56 degrees C for 30 min and 5) showed an absence of antigenicity of IgE in a partial purified inhibitor with a molecular weight of 30,000-40,000. The inhibitory effect seemed to be reversible.

Chemotactic receptor of cord blood granulocytes to the synthesized chemotactic peptide N-formyl-methionyl-leucyl-phenylalanine.

Chemotactic mobility of cord blood granulocytes (CBG) was studied under varying concentrations of synthesized chemotactic peptide, N-formyl-methionyl-leucyl-phenylalanine (f . Met-Leu-Phe). The maximal chemotactic mobility was found at a concentration of 2 X 10(-7)M in CBG and 1 X 10(-7)M in adult blood granulocytes (ABG) (n = 8). The maximal distance of granulocyte mobility of CBG was significantly shorter than that of ABG (P less than 0.01). The number of chemotactic receptors and the affinity constant were assayed by the competitive inhibition method, using f . Met-Leu-[3H]Phe, and the data were subjected to Scatchard analysis. The number of chemotactic receptors of ABG was 2.5-fold of CBG, as shown by 21,800 +/- 7800 per cell in ABG (n = 3) and 9000 in CBG (n = 1). This figure was confirmed by the one point assay method by increasing the sample numbers of cord blood. It was found that bound chemotactic peptide (X 10(-14) moles/10(7) cells) was 7.9 +/- 0.7 in ABG (n = 4) and 3.4 +/- 0.7 in CBG (n = 5). Affinity constants were similar in CBG and ABG.

Skin histidase activity and urine formiminoglutamic acid (FIGLU) in patients with histidinemia found by screening newborn infants.

Skin histidase activities and urine formiminoglutamic acid (FIGLU) levels were measured in 20 patients with histidinemia, identified by Guthrie's screening method, and their family members as well as control subjects. There was a significant positive correlation between skin histidase activities and the amounts of urine FIGLU. Although the difference of skin histidase activity and the amount of urinary FIGLU was significant between any two of the three groups (i.e. controls, parents and patients; p less than 0.005), these levels ranged widely and a considerable number of the cases overlapped among groups. When a discriminant function was computed to obtain the minimum probability of misclassification between the groups using the above two parameters, a better segregation was observed. However, even though the number of misclassifications decreased, the overlapping cases were still present, especially between the parent and patient groups. It is concluded that either skin histidase activity, urine FIGLU, or both, can be used as genetic markers of the disease to a large but still limited extent.