An insertion mutation in the Apoe gene associated with spontaneous hyperlipidemia in mice.

Introduction: Spontaneously hyperlipidemic (SHL) mice, a mouse strain derived from an inbred strain of Japanese wild (original)-type mice (KOR; Mus musculus molossinus), show high plasma cholesterol concentrations with disruption of the apolipoprotein E (Apoe) gene. However, the details of the Apoe gene of SHL mice have yet to be described. Material and methods: The DNA sequence of the Apoe gene of SHL mice was compared to that of control KOR mice in genomic DNA and cDNA analyses. Results: In the DNA analysis, a 4700-bp fragment was found to be inserted into exon 4 of the Apoe gene of SHL mice. The insertion contained two 365-bp repeats at each terminal and was flanked by a 6-bp target duplication at each side. The inserted fragment produced a frameshift of an early stop codon, resulting in a protein product that consisted of 87 amino acids in SHL mice compared to 311 amino acids in control KOR mice. Conclusions: These findings provide useful information about the molecular basis of SHL mice and related lipid disorders.

Distribution of single nucleotide polymorphisms in the CXCR1 gene and association with calf diseases in Japanese Black cattle.

The chemokine (C-X-C) receptor 1 (CXCR1) expressed on the neutrophil surfaces interacts primarily with interleukin-8 (IL-8) and has an important role in immune response. Two interesting single nucleotide polymorphisms (SNPs), SNP CXCR1+777G>C and SNP CXCR1-1768T>A, that exhibit an association with subclinical mastitis and milk quality in dairy cattle, respectively, have been reported in the bovine CXCR1 gene. The aim of this study was to demonstrate the presence of the two SNPs in the CXCR1 gene of Japanese Black cattle and examine the association between the SNPs and clinical diseases including intestinal and respiratory diseases in calves. Genotyping of the SNPs in healthy Japanese Black cattle showed that the SNPs were also present in Japanese Black cattle with gene frequencies of 0.37 and 0.15 for the C-type allele in SNP CXCR1+777 and for the A-type allele in SNP CXCR1-1768, respectively. Statistical analysis of the genotype distribution of the SNPs in the bovine CXCR1 gene in healthy and clinical intestinal or respiratory diseased Japanese Black cattle indicated no significant association of the SNPs with clinical diseases in the calves. However, a significant correlation of the number of A alleles in SNP CXCR1-1768 with white blood cell (WBC) and platelet counts was found in the disease group. It is possible that the SNP in the bovine CXCR1 gene plays a role in modulating the hematological profile of WBC and platelet counts.

Molecular sexing in Japanese murrelet (Synthliboramphus wumizusume) and a tandem-repeat polymorphism on the W chromosome.

The Japanese murrelet (Synthliboramphus wumizusume) is an endangered small seabird species in Japan. Molecular sexing using PCR targeting of the gene encoding chromodomain helicase DNA-binding protein 1(CHD1) has been used for sex identification. Specifically, PCR using any of three commonly used primer sets (CHD1F/1R, 2550F/2718R and P2/P8) has permitted sexing in many bird species. CHD1F/1R and 2550F/2718R permitted molecular sexing in Japanese murrelet; however, P2/P8 did not permit. To generate a primer pair that permits efficient molecular sexing in this species, a new primer set, CHD1F1/1R1, was prepared to permit amplification of smaller products from degraded DNA samples. The electrophoretic patterns of PCR products amplified with the new primer set were easily classified as female or male. Additionally, the PCR product indicated the presence of a polymorphism in the fragment from chromosome W. The PCR fragments of long-type (WL) and short-type (WS) polymorphisms were observed only in females. When the distribution of the CHD1 gene on chromosome W of 61 female Japanese murrelet on Biroujima Island in Miyazaki Prefecture, WL and WS were observed in 90.2% and 9.8%. The DNA polymorphism is derived from the number of copies of a 32-bp-repeat unit, with WL and WS corresponding to two and one 32-bp-repeats, respectively.

Concentrations of 137Cs radiocaesium in the organs and tissues of low-dose-exposed wild Japanese monkeys.

OBJECTIVES: Following the massive earthquake that struck eastern Japan on March 11, 2011, a large amount of radioactive material was released into the environment from the damaged reactor of the Fukushima Daiichi Nuclear Power Plant (FDNPP). After the FDNPP accident, radiocaesium was first detected in muscle samples from wild Japanese monkeys exposed to radioactive materials, and haematologic effects, changes in head size, and delayed body weight gain were also reported, but little is known about the distribution of 137Cs in the organs and tissues of wild Japanese monkeys. RESULTS: We detected the 137Cs in various organ and tissue samples of 10 wild Japanese monkeys inhabiting the forested areas of Fukushima City that were captured between July and August 2012. Among muscle, brain, heart, kidney, liver, lung, and spleen, muscle exhibited the highest and the brain the lowest 137Cs concentration. The concentration (mean ± SD) of 137Cs in muscle, brain, heart, kidney, liver, lung, and spleen was 77 ± 66, 26 ± 22, 41 ± 35, 49 ± 41, 41 ± 38, 53 ± 41, and 53 ± 51 Bq/kg, respectively. These results can help us understand the biological effects of long-term internal radiation exposure in non-human primates.

The teleost Oryzias latipes shows telomere shortening with age despite considerable telomerase activity throughout life.

Previous studies of telomeres and telomerase have focused mostly on mammals, and data for other vertebrates are limited. We analyzed both telomere length (terminal restriction fragment length) and telomerase activity in a small freshwater teleost fish, the medaka (Oryzias latipes), and found that the telomeres shorten during ageing despite the fact that a considerable amount of telomerase activity is ubiquitously detectable throughout the life of the fish. Since the telomere attrition rate during development was greater than that in adulthood, telomere length is inversely correlated with the increase in body length. The difference in telomere length among medaka individuals was similar to that in humans, and the individual specific differences were evident even at the earliest embryonic stage. Telomerase activity was ubiquitously detectable not only in the body of the embryo but also in the systemic organs of mature individuals throughout their entire life span. These data suggest that telomere attrition during ageing in medaka, which is similar to that in humans, may be a major factor determining their mortality, and that telomere maintenance through strong telomerase activity may be required for the characteristic lifelong continuous growth of this fish.

Characterization of cDNA and genomic sequences encoding a canine chemokine receptor, CXCR4 and its ligand CXCL12.

The interaction of chemokine receptor CXCR4 and its functional ligand CXCL12 plays a key role in bone marrow hematopoiesis, neuronal and cardiovascular development, and organization of the immune system. Despite the importance of the CXCL12-CXCR4 axis for regulating hematopoiesis, information on the canine CXCR4 and CXCL12 genes is insufficient. In this present study, we identified the canine counterparts of the CXCR4 and CXCL12 cDNAs and genes. The amino acid sequence encoding canine CXCR4 showed the structural characteristics of seven transmembrane domain G protein-coupled receptors and high homology with those of humans and other animals. Two isoforms, CXCL12 alpha and CXCL12 beta, were identified in dogs, as described in human and other animals. The gene structures for canine CXCR4 and CXCL12 were similar to those of other animals. The canine CXCL12 gene structure indicated that the transcripts of the isoforms arose from alternative mRNA splicing. A single nucleotide polymorphism (SNP) with synonymous substitution was observed in the exon of the canine CXCL12 gene. mRNAs encoding canine CXCR4 and CXCL12 were expressed widely and constitutively. Molecular homology and constitutive expression of CXCR4 and CXCL12 mRNAs in canine normal tissues suggests critical roles in hematopoiesis and trafficking of leukocytes, as shown in other animals.

Xanthine urolithiasis in a cat: a case report and evaluation of a candidate gene for xanthine dehydrogenase.

Xanthine urolithiasis was found in a 4-year-old spayed female Himalayan cat with a 10-month history of intermittent haematuria and dysuria. Ultrasonographs indicated the existence of several calculi in the bladder that were undetectable by survey radiographic examination. Four bladder stones were removed by cystotomy. The stones were spherical brownish-yellow and their surface was smooth and glossy. Quantitative mineral analysis showed a representative urolith to be composed of more than 95% xanthine. Ultrasonographic examination of the bladder 4.5 months postoperatively indicated the recurrence of urolithiasis. Analysis of purine concentration in urine and blood showed that the cat excreted excessive amounts of xanthine. In order to test the hypothesis that xanthinuria was caused by a homozygote of the inherited mutant allele of a gene responsible for deficiency of enzyme activity in purine degradation pathway, the allele composition of xanthine dehydrogenase (XDH) gene (one of the candidate genes for hereditary xanthinuria) was evaluated. The cat with xanthinuria was a heterozygote of the polymorphism. A single nucleotide polymorphism analysis of the cat XDH gene strongly indicated that the XDH gene of the patient cat was composed of two kinds of alleles and ruled out the hypothesis that the cat inherited the same recessive XDH allele suggesting no activity from a single ancestor.

Telomere attrition and restoration in the normal teleost Oryzias latipes are linked to growth rate and telomerase activity at each life stage.

Telomere shortening occurs when cells divide, both in vitro and in vivo. On the other hand, telomerase is able to maintain telomere length in cells by adding TTAGGG repeats to the ends of telomeres. However, the interrelationships existing among telomere length, telomerase activity and growth in vertebrates remain to be clarified. In the present study we measured telomere length (terminal restriction fragment length), telomerase activity and body growth of Oryzias latipes from the embryo stage until senescence. During the rapid growth stage (age 0-7 months), telomeres shortened in parallel with decreasing telomerase activity. Then, during adolescence (age 7 months - 1 year), telomeres lengthened quickly as growth slowed and telomerase activity increased. In the adult stage (age 1-4 years) characterized by little growth, telomerase activity decreased gradually and telomeres shortened. Our data indicate that telomere attrition and restoration are linked to growth and telomerase activity, and suggest that critical loss of telomere homeostasis is associated with mortality in this animal.

Molecular Characterization of the Cytidine Monophosphate-N-Acetylneuraminic Acid Hydroxylase (CMAH) Gene Associated with the Feline AB Blood Group System.

Cat's AB blood group system (blood types A, B, and AB) is of major importance in feline transfusion medicine. Type A and type B antigens are Neu5Gc and Neu5Ac, respectively, and the enzyme CMAH participating in the synthesis of Neu5Gc from Neu5Ac is associated with this cat blood group system. Rare type AB erythrocytes express both Neu5Gc and Neu5Ac. Cat serum contains naturally occurring antibodies against antigens occurring in the other blood types. To understand the molecular genetic basis of this blood group system, we investigated the distribution of AB blood group antigens, CMAH gene structure, mutation, diplotypes, and haplotypes of the cat CMAH genes. Blood-typing revealed that 734 of the cats analyzed type A (95.1%), 38 cats were type B (4.9%), and none were type AB. A family of three Ragdoll cats including two type AB cats and one type A was also used in this study. CMAH sequence analyses showed that the CMAH protein was generated from two mRNA isoforms differing in exon 1. Analyses of the nucleotide sequences of the 16 exons including the coding region of CMAH examined in the 34 type B cats and in the family of type AB cats carried the CMAH variants, and revealed multiple novel diplotypes comprising several polymorphisms. Haplotype inference, which was focused on non-synonymous SNPs revealed that eight haplotypes carried one to four mutations in CMAH, and all cats with type B (n = 34) and AB (n = 2) blood carried two alleles derived from the mutated CMAH gene. These results suggested that double haploids selected from multiple recessive alleles in the cat CMAH loci were highly associated with the expression of the Neu5Ac on erythrocyte membrane in types B and AB of the feline AB blood group system.

Characterization of Kdap, a protein secreted by keratinocytes.

Using a signal sequence-trap we identified a human gene encoding a polypeptide of 99 amino acids with a putative signal sequence. The gene was identical to keratinocyte differentiation-associated protein (Kdap), which was reported previously by Oomizu et al (Gene 256: 19-27, 2000) to be expressed in embryonal rat epidermis at the mRNA level. In humans, we found Kdap mRNA expression to be restricted to epithelial tissue at high levels. The 12.5 kDa protein was detected in culture supernatant of keratinocytes and those transfected adenovirally with the Kdap gene. In normal skin, Kdap protein was found exclusively within lamellar granules of granular keratinocytes and in the intercellular space of the stratum corneum. By contrast, in lesional skin of patients with psoriasis, Kdap was expressed more widely throughout suprabasal keratinocytes. When induced to differentiate in vitro, keratinocytes showed marked upregulation of Kdap mRNA expression similar to that of involucrin mRNA, but with differing kinetics. Finally, a spliced variant of Kdap mRNA was generated by alternative splicing mechanisms. Our studies indicate that human Kdap resembles rat Kdap with respect to tissue and cell expression at the mRNA level and that Kdap is a low-molecular-weight protein secreted by keratinocytes. Thus Kdap may serve as a soluble regulator of keratinocyte differentiation.

Molecular analysis of the human esterase D gene ESD(*)Q0(yonago) responsible for incompatibility in a Japanese paternity case.

In a Japanese paternity test, an alleged father was excluded only by reverse homozygosity of esterase D (ESD) phenotypes (mother, ESD 1; child, ESD 1; alleged father, ESD 2) out of 43 classical and DNA markers investigated. To solve the aberrant inheritance of the ESD phenotypes observed between them, fragments for all eight coding exons amplified by polymerase chain reaction (PCR) were subjected to DNA analysis. The child and alleged father shared a null allele, originating from ESD(*)1. It was characterized by having TGA for the stop codon instead of TCA for serine at codon 63. Thus, the sharing of a rare null gene, ESD(*)Q0(yonago), increased the probability of paternity.

Evaluation of intestinal intramucosal pH, arterial and portal venous blood gas values, and intestinal blood flow during small intestinal ischemia and reperfusion in dogs.

OBJECTIVES: To determine whether small intestinal ischemia and reperfusion affects intestinal intramucosal pH (pHi), arterial and portal venous blood gas values, and intestinal blood flow (IBF) and to investigate relationships between regional intestinal tissue oxygenation and systemic variables in dogs. ANIMALS: 15 healthy adult Beagles. PROCEDURE: Occlusion of superior mesenteric artery (SMA) for 0, 30, or 60 minutes, followed by reperfusion for 180 minutes, was performed; IBF, pHi, arterial and portal venous blood gas values, arterial pressure, and heart rate were measured at various time points; and intestinal mucosal injury was histologically graded. RESULTS: Occlusion of the SMA induced significant decreases in pHi and IBF. After the release of the occlusion, IBF returned rapidly to baseline values, but improvement in pHi was slow. Arterial and portal venous blood gas analyses were less sensitive than tonometric measurements of pHi, and there was no correlation between results of blood gas analyses and tonometric measurements. Histologic score for intestinal mucosal injury increased significantly, depending on duration of ischemia, and there was a correlation between tonometric results and the histologic score. CONCLUSIONS AND CLINICAL RELEVANCE: Results suggest that it is difficult to accurately evaluate local oxygenation disorders by monitoring at the systemic level, whereas clinically pHi is the only reliable indicator of inadequate regional intestinal tissue oxygenation in dogs.

Kinetics of endotoxin concentration and tumor necrosis factor-alpha, interleukin-1beta, and interleukin-6 activities in the systemic and portal circulation during small intestinal ischemia and reperfusion in dogs.

OBJECTIVE: To determine whether small intestinal ischemia and reperfusion induces bacterial translocation and proinflammatory cytokine response in either the systemic or portal circulation in dogs. ANIMALS: 17 healthy adult Beagles. PROCEDURE: The superior mesenteric artery (SMA) was occluded for 0 (group-3 dogs), 30 (group-1 dogs), or 60 (group-2 dogs) minutes, followed by reperfusion for 180 minutes; serum lactate and endotoxin concentrations and tumor necrosis factor-alpha (TNF-alpha), interleukin-1beta (IL-1beta), and IL-6 activities in the systemic and portal circulation and intramucosal pH were measured at various time points. RESULTS: In group-2 dogs, TNFalpha activity was found to be significantly increased in the portal circulation, peaking at 60 minutes of reperfusion; TNF-alpha activity, in the systemic circulation, gradually increased from 60 minutes of reperfusion to the end of the experiment; however, the increase was not significant. In group-1 and -2 dogs, IL-6 activities significantly and gradually increased in the systemic and portal circulation during the reperfusion phase, and the magnitude of these increases was dependent on the duration of the ischemic phase. There were no significant changes in IL-1beta activity or endotoxin concentration in any dog group. CONCLUSIONS AND CLINICAL RELEVANCE: Results of the our study indicate that intestinal ischemia and reperfusion leads to significant increases of the circulating TNF-alpha and IL-6 activities, depending on the duration of the ischemia phase, in the absence of detectable endotoxin in the circulation. This finding suggests that intestinal ischemia and reperfusion induces a systemic proinflammatory cytokine response in dogs.

The genetic association study between polymorphisms in uncoupling protein 2 and uncoupling protein 3 and metabolic data in dogs.

BACKGROUND: The uncoupling proteins (UCPs) in the mitochondrial inner membrane are members of the mitochondrial anion carrier protein family that play an important role in energy homeostasis. Genetic association studies have shown that human UCP2 and UCP3 variants (SNPs and indels) are associated with obesity, insulin resistance, type 2 diabetes mellitus, and metabolic syndrome. The aim of this study was to examine the genetic association between polymorphisms in UCP2 and UCP3 and metabolic data in dogs. RESULTS: We identified 10 SNPs (9 intronic and 1 exonic) and 4 indels (intronic) in UCP2, and 13 SNPs (11 intronic and 2 exonic) and one indel (exonic) in UCP3, by DNA sequence analysis of 11 different dog breeds (n=119). An association study between these UCP2 and UCP3 variants and the biochemical parameters of glucose, total cholesterol, lactate dehydrogenase and triglyceride in Labrador Retrievers (n=50) showed that none of the UCP2 polymorphisms were significantly associated with the levels of these parameters. However, four UCP3 SNPs (intron 1) were significantly associated with total cholesterol levels. In addition, the allele frequencies of two of the four SNPs associated with higher total cholesterol levels in a breed that is susceptible to hypercholesterolemia (Shetland Sheepdogs, n=30), compared with the control breed (Shiba, n=30). CONCLUSION: The results obtained from a limited number of individuals suggest that the UCP3 gene in dogs may be associated with total cholesterol levels. The examination of larger sample sizes and further analysis will lead to increased precision of these results.

Low blood cell counts in wild Japanese monkeys after the Fukushima Daiichi nuclear disaster.

In April 2012 we carried out a 1-year hematological study on a population of wild Japanese monkeys inhabiting the forest area of Fukushima City. This area is located 70 km from the Fukushima Daiichi Nuclear Power Plant (NPP), which released a large amount of radioactive material into the environment following the Great East Japan Earthquake of 2011. For comparison, we examined monkeys inhabiting the Shimokita Peninsula in Aomori Prefecture, located approximately 400 km from the NPP. Total muscle cesium concentration in Fukushima monkeys was in the range of 78-1778 Bq/kg, whereas the level of cesium was below the detection limit in all Shimokita monkeys. Compared with Shimokita monkeys, Fukushima monkeys had significantly low white and red blood cell counts, hemoglobin, and hematocrit, and the white blood cell count in immature monkeys showed a significant negative correlation with muscle cesium concentration. These results suggest that the exposure to some form of radioactive material contributed to hematological changes in Fukushima monkeys.

Concentration of radiocesium in the wild Japanese monkey (Macaca fuscata) over the first 15 months after the Fukushima Daiichi nuclear disaster.

Following the massive earthquake that struck eastern Japan on March 11, 2011, a nuclear reactor core meltdown occurred at the Fukushima Daiichi Nuclear Power Plant, operated by Tokyo Electric Power Company, and was followed by the release of large amounts of radioactive materials. The objective of this study was to measure the concentration of radiocesium (134)Cs and (137)Cs in the muscle of Japanese monkeys (Macaca fuscata) inhabiting the forest area of Fukushima City and to determine the change in concentration over time as well as the relationship with the level of soil contamination. Cesium concentrations in the muscle of monkeys captured at locations with 100,000-300,000 Bq/m(2) were 6,000-25,000 Bq/kg in April 2011 and decreased over 3 months to around 1,000 Bq/kg. However, the concentration increased again to 2,000-3,000 Bq/kg in some animals during and after December 2011 before returning to 1,000 Bq/kg in April 2012, after which it remained relatively constant. This pattern of change in muscle radiocesium concentration was similar to that of the change in radiocesium concentration in atmospheric fallout. Moreover, the monkeys feed on winter buds and the cambium layer of tree bark potentially containing higher concentrations of radiocesium than that in the diet during the rest of the year. The muscle radiocesium concentration in the monkeys related significantly with the level of soil contamination at the capture locations.

Analysis of mitochondrial DNA HVR1 haplotype of pure-bred domestic dogs in Japan.

To develop DNA markers for forensic analysis, we examined the hypervariable region 1 (HVR1) sequences of 447 pure-bred domestic dogs (Canis lupus familiaris) that had been bred and raised in Japan. HVR1 is a 660-bp stretch of mitochondrial (mt) DNA. Among the 447 HVR1 sequences examined, we identified 58 haplotypes from 47 single nucleotide polymorphisms (SNPs) and two insertion-deletion (InDel) polymorphisms. The haplotype diversity inferred from inter-breed analysis (N=154, 88 breeds) was 0.929±0.011. Intra-breed analysis showed that the haplotype diversity of Golden Retrievers (N=53), Labrador Retrievers (N=67), Miniature Dachshunds (N=61), Toy Poodles (N=62), and Welsh Corgis (N=50) was 0.624±0.052, 0.722±0.029, 0.922±0.010, 0.877±0.020, and 0.443±0.084, respectively. The results of this genotype analysis were used to construct a dataset consisting of dog mtDNA HVR1 sequences for use in forensic applications in Japan.

Identification of microphthalmia-associated transcription factor isoforms in dogs.

Microphthalmia-associated transcription factor (MITF) belongs to the basic helix-loop-helix leucine zipper (bHLH-LZ) family of transcription factors, and plays an important role in the development and differentiation of melanocytes, mast cells, and osteoclasts. To investigate canine MITF isoforms, cDNA cloning with 5'-rapid amplification of cDNA ends was performed. Three canine isoforms, the counterparts of human MITF-M, MITF-H, and MITF-A, were determined. Canine MITF-M, MITF-H, and MITF-A were highly homologous to the nucleotide sequences in isoform-specific 1M, 1H, and 1A with 100%, 98%, and 97% identity in humans, respectively. When scanning of the canine genome was performed to identify the homologous regions to human MITF isoforms, in addition to MITF-M, MITF-H, and MITF-A isoforms, the homologous sequences to all the other isoforms observed in humans existed in the canine genome sequence of chromosome 20. Comparison of the homologous sequences for MITF isoforms among dogs and humans suggested that MITF-D, MITF-E and MITF-J may be expressed in canine adult tissues in addition to canine MITF-M, MITF-H, and MITF-A. However, RT-PCR showed that canine MITF-E and MITF-J, but not canine MITF-D were expressed. Although the mRNAs encoding canine MITF-H and MITF-A were expressed widely, MITF-M, MITF-E and MITF-J were expressed in tissue-specific patterns. Two types of mRNAs with and without an 18-base insert were detected for five MITF isoforms in canine tissues. MITF-M distal enhancer (MDE) was also located upstream from canine exon 1M.

Molecular basis of ESD*5 and ESD*7 and haplotype analysis with new polymorphisms in introns.

The two polymorphic alleles of esterase D (ESD), ESD*5 and ESD*7, are specific to Europeans and Asians, respectively. In this study the molecular basis was characterized: ESD*5, arising from ESD*1, has a G to A transition, resulting in Gly257(GGT) --> Asp(GAT); and ESD*7, originating from ESD*2, has an A to G transition, resulting in Asp231(GAT) --> Gly(GGT). Glycine is also involved in the common ESD*1/ESD*2 polymorphism [Gly190(GGA) --> Glu(GAA)]. Haplotype analysis using a few novel intragenic polymorphisms showed strong associations among polymorphic sites, suggesting that recombination has been less frequent in the human ESD gene, although it spans about 25 kb from exon 1 to exon 10. A marked difference was observed in the distribution of haplotype frequencies between Germans and Japanese.

Apoptotic lymphocytes induced by surgical trauma in dogs.

PURPOSE: An investigation was made of the population and function of lymphocytes in canine peripheral blood, in animals with or without laparotomy under inhalation anesthesia. METHODS: Fourteen healthy beagles were allocated to two experimental groups: laparotomy (group A) and without laparotomy (group B). Induction of anesthesia in both groups was carried out with an intravenous injection of thiopentone, and was maintained by isoflurane inhalation. Ten blood samples, consisting of 10 ml of venous blood withdrawn by venipuncture into syringes containing 200 units of preservative-free heparin, were taken, from before anesthesia to the 28th postoperative day. The blood samples were collected, and the number of lymphocytes, the lymphocyte subpopulations, the proportion of apoptotic lymphocytes, and plasma cortisol level were measured, and the blastoid transformation of lymphocytes was observed. RESULTS: Lymphopenia was observed in both groups after anesthesia. Flow cytometry indicated a greater reduction in the proportion of T lymphocytes than of B lymphocytes. Blast transformation was also depressed in both groups. Progression of apoptosis after anesthesia was demonstrated in both groups, with a higher percentage of apoptotic cells being observed in group A at 12 h after anesthesia (28.5 +/- 3.2% by TUNEL assay). Plasma levels of cortisol were elevated to a greater extent in group A at the end of anesthesia (10.3 +/- 0.8 microg/dl) than in group B (7.8 +/- 1.9 microg/dl). CONCLUSIONS: These results indicate that surgical trauma concomitant with anesthesia could impair immunocompetence by reducing the number and function of lymphocytes.