Construction and validation of a scoring system to predict resistance to chemotherapeutic drugs using gene expression profiles in canine lymphoma.

The present study aimed to verify the changes in the expression levels of 13 candidate genes associated with chemotherapy resistance and to construct a scoring system to predict resistance to these drugs. The expression levels of the 13 candidate genes were compared between 20 dogs with lymphoma that were sensitive to drugs used in CHOP-based protocol and 16 dogs with lymphoma that were resistant to these drugs. The expression levels of six genes; ASNS, CCR3, CALCA, FCER1A, LOC448801, and EDNRB were significantly different between the two groups. A scoring system to predict resistance to cyclophosphamide, doxorubicin and vincristine, which are used in CHOP-based protocol, was constructed based on expression levels of the six genes in these 36 dogs using logistic regression models. After internal validation, sensitivity and specificity of the scoring system were 0.759 and 0.853, respectively. External validation was conducted in another cohort of 33 dogs with lymphoma, and sensitivity and specificity of the scoring system were 0.800 and 0.696, respectively. In conclusion, this study identified six genes associated with resistance to drugs used in CHOP-based protocol in canine lymphoma and proposed a novel scoring system to predict resistance to these drugs. This system might be beneficial in selecting the most appropriate chemotherapy protocol for individual dogs with lymphoma.

Development of canine X-chromosome inactivation pattern analysis for the detection of cell clonality by incorporating the examination of the SLIT and NTRK-like family member 4 (SLITRK4) gene.

X-chromosome inactivation pattern (XCIP) analysis can be used to assess the clonality of cell populations of various origin by distinguishing the methylated X chromosome from the unmethylated X chromosome. In this study, the utility of XCIP analysis was improved by incorporating the examination of AC dinucleotide repeats in SLIT and NTRK-like family member 4 (SLITRK4) gene into the previously reported CAG repeat examination of androgen receptor (AR) gene in dogs. The rate of heterozygosity when both genes were analysed (125/150, 83.3%) was higher than AR gene examination alone (86/150, 57.3%). Blood samples from heterozygous dogs in either AC-1 or AC-2 of SLITRK4 gene were examined for the corrected inactivation allele ratio (CIAR), resulting in the determination of a reference range of CIAR <3.8 in non-neoplastic cell/tissue samples. Using this analytical method, 49% (21/43) of neoplastic tissue samples from dogs showed a CIAR >3.8, indicating the presence of a clonal population. Through the present study, the availability of canine XCIP analysis was improved by incorporating the examination of the SLITRK4 gene, providing a highly useful laboratory examination system for the detection of the clonality of various cell/tissue samples in dogs.

Clinical and clinicopathological characteristics of acute lymphoblastic leukaemia in six cats.

OBJECTIVES: To investigate the clinical characteristics of feline acute lymphoblastic leukaemia patients diagnosed according to the recent diagnostic criteria for the equivalent canine condition. MATERIALS AND METHODS: The medical records of six cats diagnosed with acute lymphoblastic leukaemia were retrospectively reviewed to extract data on clinicopathological characteristics and outcomes. The lymphoid origin of the tumour cells was confirmed by polymerase chain reaction for antigen receptor gene rearrangement, flow cytometry or immunohistochemistry. RESULTS: Non-specific clinical signs such as lethargy and anorexia were common, and anaemia and thrombocytopenia were also commonly identified. Leucocytosis was observed in four cats and leucopenia was observed in two; the number of lymphoblasts in the peripheral blood samples varied among the cases. Lymphoblasts originated from B-cell lineage in four cats and T-cell lineage in one, and those of another cat were positive for both B-cell marker CD21 and T-cell marker CD8. Five of the six cats were treated with cytotoxic chemotherapy, and a partial response was obtained in two. The median overall survival was 55 days (range: 1 to 115). CLINICAL SIGNIFICANCE: Acute lymphoblastic leukaemia should be considered if lymphoblasts are observed in peripheral blood, even if their number is small. The prognosis for cats that have acute lymphoblastic leukaemia is as poor as that for dogs, and further studies are needed to develop effective treatment.

Cross-antigenicity among EV71 strains from different genogroups isolated in Yamagata, Japan, between 1990 and 2007.

We isolated and identified six subgenogroups (B2, B4, B5, C1, C2, and C4) of enterovirus 71 (EV71) between 1990 and 2007 in Yamagata, Japan. We measured neutralizing antibody (NT Ab) titers against those subgenogroup strains and the BrCr reference strain for antigenic analysis. Serological analysis of 83 residents in Yamagata in 2004 showed that differences in the NT Ab titer of each individual against the different subgenogroups were mostly within 4-fold. Furthermore, sera from guinea pigs, immunized with the B2 and C1 strains indicated cross-antigenicity among the seven different subgenogroups. In conclusion, our results showed that cross-antigenicity exists among EV71 strains from different subgenogroups circulating in the community through genomic evolution. Our results also suggest that eliciting neutralizing antibodies against one genotype is likely to confer cross-neutralization against other genotypes.

[The influence of ulinastatin on renal function during anesthesia for renal transplantation].

The purpose of this study was to evaluate the influence of ulinastatin (UST) on renal function and changes in granulocyte elastase during anesthesia in 10 living related and 10 cadaveric renal transplantations. UST (300,000 units) was administered after the induction of anesthesia and renal vessel anastomosis, respectively. The living patients who received UST had greater urine volume during the operative and postoperative periods. In addition, the granulocyte elastase values of these patients showed relative suppressive changes during the intraoperative and postoperative periods. In cadaveric renal transplant patients, UST did not influence serum BUN or creatinine, although urine volume was greater in the UST-treated patients than in the patients who did not receive. We could not conclude that the intraoperative administration of UST affected the renal function of renal transplants, but we did find that UST is useful to suppress granulocyte elastase which is concerned with the index of stress or inflammation. It is necessary to investigate further the method of administration of UST for renal transplantation.

[Use of vecuronium bromide for anesthesia for renal transplantation].

We used vecuronium on 7 patients for renal transplants and recorded the muscle relaxant effect using an Anesthesia and Brain Monitor; we also compared them with patients with normal renal function. Results show that there were no significant differences between renal transplant patients and patients with normal renal functions in regard to the time of appearance of effect, duration of effect and recovery rate but the effect was somewhat prolonged in the renal transplant patients. Therefore, although vecuronium is said to be a muscle relaxant with little renal dependency, and it is used on renal transplant patients with renal insufficiency, the possibility of a prolonged effect must be considered. It is important to monitor continuously by a muscle relaxant monitor, grasp adequately the conditions of muscle relaxation, and observe carefully the postoperative respiratory conditions.

[Changes in catecholamine and immunoglobulin during anesthesia for cardiac surgery--a comparison of valve replacement and aorto-coronary bypass surgery].

During a period of extracorporeal circulation, which represents a massive invasion for the body, the patient is in a state of controlled shock. In this study, we examined changes in catecholamines and immunoglobulins in patients grouped according to whether they underwent valve replacement or aorto-coronary bypass surgery. Anesthesia was induced with fentanyl, and cardiopulmonary bypass in all patients was carried out using a bubble-type heart-lung machine. The epinephrine and norepinephrine levels increased during the procedures, but those patients having aorto-coronary bypass surgery showed milder rises than valve replacement patients. IgA, IgG and IgM all dropped as well, but the change in IgG was the greatest, and the decreases were more marked in valve replacement patients than in aorto-coronary bypass patients. It was concluded that, since reactions were different according to the disorder present and were reflected in the variations in catecholamine and immunoglobulin levels, great care is necessary when the patient is removed from the heart-lung machine.

[Hemodynamic changes before and after anesthesia for renal transplantation].

Renal transplantation has come into use as a treatment for renal insufficiency, but infusion management before and after anesthesia for this operation is important. In order to prevent acute necrosis of the uriniferous tubules and to obtain urine outflow in early postoperative stage, a recent practice has been to give rapid infusions of large amounts of fluid, starting during the anastomosis of the renal vessels. We gave a large amount of intraoperative fluid to six patients undergoing transplantation of cadaver kidneys. A Swan-Ganz catheter was inserted into the pulmonary artery and infusion management was performed so as to maintain the pulmonary arterial pressure above 15 mmHg and the pulmonary capillary wedge pressure above 10 mmHg. The cardiac output increased as a result, and no pulmonary edema was seen. We believe that our method of infusion management using a Swan-Ganz catheter is a useful technique in such cases.

A microplate method for isolation of viruses from infants and children with acute respiratory infections.

Between December 1984 and December 1986, a microplate technique was adopted for isolation of viruses from infants and children with acute respiratory infections. By using two kinds of tissue culture microplates, i.e., the HHVM plate, containing human embryonic fibroblast (HEF), HEp-2, Vero and MDCK cells, and the MK plate which contains secondary monkey kidney cells, 1,080 field viruses were isolated from 1,061 (24.9%) out of 4,254 throat swabs. Of these 1,080 isolates, 1,003 (92.9%) were recovered in the HHVM plates and the remaining 77 (7.1%) in the MK plates. With the HHVM plate, influenza A and B viruses were cultivated in MDCK, RS virus in HEp-2, parainfluenza and mumps viruses in Vero, adenoviruses in both HEF and HEp-2, polioviruses in HEF, HEp-2 and Vero, coxsackie B viruses in both HEp-2 and Vero, rhino and echo viruses in HEF, herpes simplex virus in both HEF and HEp-2, and cytomegalovirus in HEF, although MK were more sensitive than Vero to parainfluenza and coxsackie B viruses. There was no difference in the rate of isolation of viruses between the microplate and ordinary tube methods. Cross contamination in the microplates was negligible for routine work.

[Observations on mortality in selected working populations].

Mortality among employees of selected enterprises in which excellent health care programmes are carried on, was observed to evaluate employees' health levels and to establish objectives for future health care programmes. Also, these data were compared with those for all Japanese and for Japanese employees belonging to the corresponding occupational groups. Seventy-three enterprises which continuously participated in the mortality survey implemented by the Japan Research Organization of Industrial Health Care for the period between 1976 and 1980, were subjected to this study. The observed employees amounted to 3,502,580 person-years. Sex and age distributions of the employees were available for 47 enterprises and the observed employees totalled 2,598,672 person-years. The main results are as follows: Mortality rate (per 100,000) is stable between 140 and 150 throughout the observed period and the average value is 145.0. The average mortality rate for males, aged 40 to 54, is 272.6. Malignant neoplasms were the main cause of death and account for 37-38%. In second place and below are heart diseases, cerebrovascular diseases, accidents, suicide and liver cirrhosis. Malignant neoplasms, especially of the stomach, lung and pancreas, show a trend to increase, and cerebrovascular diseases and liver cirrhosis show a trend to decrease. Among heart diseases, ischemic heart disease accounts for about 40% and shows no marked fluctuation. Among cerebrovascular diseases, the relative frequency of subarachnoid hemorrhage is increasing. Comparing the mortality rates for males aged 40 to 54 by industry, "Iron, steel & nonferrous metal manufacturing" and "Electricity and gas supply" show significantly higher values, and "Finance & insurance" and "Communications" show significantly lower values than the total. The distribution of main causes of death for males, aged 40 to 54, was compared by major occupational groups using Proportional Mortality Ratio (PMR). A significantly high frequency of malignant neoplasms (especially of the stomach) is observed for "Professional & technical workers, managers and officials" and that of suicide for "Craftmen, production process workers and labourers" and that of cerebrovascular diseases (especially cerebral hemorrhage) for "Protective service workers." Standardized Mortality Ratio (SMR) for males, aged 20 to 54, from all causes of death, calculated on the basis of all Japanese males in 1978 is 0.57. SMR for this population from malignant neoplasms is 0.89.(ABSTRACT TRUNCATED AT 400 WORDS)