GM2 gangliosidosis variant 0 (Sandhoff-like disease) in a family of toy poodles.

BACKGROUND: GM2 gangliosidosis variant 0 (human Sandhoff disease) is a lysosomal storage disorder caused by deficiencies of acid ß-hexosaminidase (Hex) A and Hex B because of an abnormality of the ß-subunit, a common component in these enzyme molecules, which is coded by the HEXB gene. OBJECTIVE: To describe the clinical, pathological, biochemical, and magnetic resonance imaging (MRI) findings of Sandhoff-like disease identified in a family of Toy Poodles. ANIMALS: Three red-haired Toy Poodles demonstrated clinical signs including motor disorders and tremor starting between 9 and 12 months of age. The animals finally died of neurological deterioration between 18 and 23 months of age. There were some lymphocytes with abnormal cytoplasmic vacuoles detected. METHODS: Observational case study. RESULTS: The common MRI finding was diffuse T2-hyperintensity of the subcortical white matter in the cerebrum. Bilateral T2-hyperintensity and T1-hypointensity in the nucleus caudatus, and atrophic findings of the cerebrum and cerebellum, were observed in a dog in the late stage. Histopathologically, swollen neurons with pale to eosinophilic granular materials in the cytoplasm were observed throughout the central nervous system. Biochemically, GM2 ganglioside had accumulated in the brain, and Hex A and Hex B were deficient in the brain and liver. Pedigree analysis demonstrated that the 3 affected dogs were from the same family line. CONCLUSIONS AND CLINICAL IMPORTANCE: The Sandhoff-like disease observed in this family of Toy Poodles is the 2nd occurrence of the canine form of this disease and the 1st report of its identification in a family of dogs.

Expression of Peroxisome Proliferator-activated Receptor-γ in the Kidneys of Cats with Chronic Kidney Disease.

Peroxisome proliferator-activated receptor (PPAR)-γ plays an important role in various cellular functions and its activation exerts protective effects in kidney diseases. In the present study, chronic kidney disease in cats was examined, and changes in renal expression of PPARγ were observed by use of immunohistochemistry. In normal kidneys, nuclei of the superficial cortical tubules, medullary tubules and glomerular cells expressed PPARγ. The vascular walls (tunica media) also showed positive expression. In diseased kidneys, the expression of PPARγ varied between the cases. Some cases showed strong expression, while others had weak expression. PPARγ expression in the nuclei of infiltrating mononuclear cells was also detected in over half of the cases. Although there was no significant relationship between the expression of renal PPARγ and the severity of kidney disease, the fact that there were many cases where the expression of renal PPARγ was reduced was an important finding, and might be one of the possible mechanisms underlying feline chronic kidney diseases.

Characterization of bradykinin-induced endothelium-independent contraction in equine basilar artery.

We investigated the effect of bradykinin (BK) on isolated equine basilar arterial rings with and without endothelium. BK induced concentration-dependent contraction of resting arterial rings and no relaxation when the rings were precontracted by prostaglandin F(2alpha). The maximal response and pD(2) value were 161.2 +/- 28.1% (to 60 mm KCl-induced contraction) and 8.24 +/- 0.25 respectively. The cumulative concentration-response curve for BK was not shifted to the right by des-Arg(9)-[Leu(8)]-BK (a B(1)-receptor antagonist), HOE140 (a B(2)-receptor antagonist) or NPC567 (another B(2)-receptor antagonist). In four of six basilar arteries, NPC567 induced concentration-dependent contraction. Indomethacin (a cyclooxygenase inhibitor), nordihydroguaiaretic acid (a lipoxygenase inhibitor), quinacrine (a phospholipase A(2) inhibitor), tetrodotoxin (a selective blocker of Na(+) channels), guanethidine (a nor-adrenergic neuron blocking drug), phentolamine (an alpha-adrenoceptor antagonist), Nomega-nitro-L-arginine (L-NNA, a nitric oxide (NO) synthase inhibitor) and endothelial denudation did not affect the BK-induced contraction. L-NNA and indomethacin induced contraction and relaxation under resting vascular tone respectively. These results suggest that endothelial cells are not involved in BK-induced contraction and that the contraction is not mediated via activation of known B(1) and B(2) receptors. Arachidonic acid metabolites and neurotransmitters like norepinephrine and NO might not play a role in BK-induced contraction in equine basilar artery.

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.

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.

Molecular prevalence of multiple genetic disorders in Border collies in Japan and recommendations for genetic counselling.

Reproductive management is necessary to prevent deleterious genetic disorders in purebred dogs, but comprehensive studies aimed at prevention of multiple underlying genetic disorders in a single breed have not been performed. The aims of this study were to examine mutant allele frequencies associated with multiple genetic disorders, using Border collies as a representative breed, and to make recommendations for prevention of the disorders. Genotyping of known mutations associated with seven recessive genetic disorders was performed using PCR assays. More than half (56%) of the Border collies had no mutant alleles associated with any of the seven disorders, suggesting that these disorders can be removed from the population over several generations. Since frequencies of each mutant allele differed among disorders, reproductive management should be performed after the establishment of prevention schemes that are appropriate for each disorder, the type and specificity of genetic test available, and the effective population size in each breeding colony.

A Practical Technique for Electron Microscopy of Buffy Coats in Dogs and Cats.

Plastic hematocrit tubes (PHTs) are convenient tools for electron microscopy (EM) of peripheral blood buffy coats, and the PHT-EM technique is expected to be a practical method for veterinary clinical medicine. In this study, fixatives composed of various concentrations of sucrose, glutaraldehyde, and phosphate buffer (PB) were tested for preparing canine and feline buffy coats. The highest quality images were obtained using a fixative consisting of 2.5% glutaraldehyde in 0.1 m PB, and it was concluded that this method allows clinicians who are inexperienced in histological techniques can conveniently transport buffy coat samples to diagnostic laboratories for analysis by EM.

Lectin-histochemical and -cytochemical study of periodic acid Schiff-positive lysosome granules as a histological feature of the female mouse kidney.

Renal proximal straight tubules (PST) of the female mouse contain periodic acid Schiff-positive lysosome granules. An excellent example of this is found in the kidneys of female DBA/2Cr mice. In the present study, lectin-histochemistry showed that lectin-positive granules occur in the PST of DBA/2Cr mice. Out of twenty-one lectins studied, the granules bound WGA, s-WGA, LEL, STL, DSL, GSL-II, VVL, RCA-I, ECL, PSA, LCA and PHA-E. Such granules were also observed in the proximal convoluted tubules (PCT). In addition, heterogeneous binding to the SBA or DBA was observed in the PST. Lectin-cytochemistry for s-WGA, STL, VVL, RCA-I, ECL and PSA, showed that: 1) lysosomes bind a higher level of s-WGA or STL than VVL, RCA-I, ECL or PSA; 2) PSA binding is similar in PST and PCT; 3) there are many PCT lysosomes that are negative for s-WGA, STL, VVL, RCA-I, and ECL lectin binding; and 4) s-WGA binding is highly specific to the lysosomes of the PST. Based on the binding specificities of each lectin, it was suggested that the mannose content of PST and PCT lysosomes is similar, and that PST lysosomes have a high level of N-acetylglucosamine, N-acetylgalactosamine, galactose or galactosyl (beta 1, 4) N-acetylglucosamine.

Sex- and strain-dependent histological features of the proximal convoluted tubular epithelium of mouse kidney: association with lysosomes containing apolipoprotein B.

In the present study, we performed comparative histological observations of ICR, BALB/c, C57BL/6, C3H/HeN and DBA/2 mice kidneys. Sex and strain differences were observed in the appearance of vacuolar structures of the proximal convoluted tubules (toluidine blue-positive granules in osmium-postfixed epoxy-resin sections). These features were especially remarkable in male DBA/2 mice. The vacuolar structures in male DBA/2 mice showed heterogeneous staining with Sudan B in frozen sections and appeared under an electron microscope as multilammelar giant dense bodies. In addition, these dense bodies showed heterogeneous acid phosphatase reactions. Immunohistochemical analyses of these structures for apolipoprotein B showed strong positive reactions. These results suggested that vacuolar structures in the proximal convoluted tubules, which were remarkable in male DBA/2 mice, were giant lysosomes containing apolipoprotein B.

Pathophysiology of brain swelling after acute experimental brain compression and decompression.

Global ischemia was created by controlled expansion of an epidural balloon for 25 minutes in Group A (six cats) and for 5 minutes in Group B (six cats). The alterations of intracranial pressure, arteriovenous oxygen content difference, cerebral metabolic rate of oxygen, cerebral blood flow, and electroencephalogram were observed until brain death or 24 hours' survival with normal intracranial pressure. The animals were then killed for brain histological examination. In four other cats, a 2% solution of Evans blue dye (4 mg/kg) was injected intravenously--immediately after deflation--resulting in 25 minutes of global ischemia. Two other cats received 5 minutes of global ischemia. The cats were killed 1 hour later. Abrupt swelling occurred in Group A, and no swelling was found in Group B. A transient absolute hyperemia was found immediately after deflation in both groups. The cerebral blood flow and cerebral metabolic rate of oxygen decreased markedly with low arteriovenous oxygen content difference and flat electroencephalogram in Group A, compared with gradual recovery of cerebral blood flow and cerebral metabolic rate of oxygen with high arteriovenous oxygen content difference and reappearance of electroencephalogram activity in Group B. The extravasation of Evans blue was observed on the compressed cerebral hemisphere, thalamus, hypothalamus, and brain stem in swelling animals and only on the compressed hemisphere in nonswelling animals. Histologically, the damage and congestive dilation of capillary, degeneration, and necrosis of neuronal and glial cell were found prominently on the hypothalamus and brain stem in the swelling group.(ABSTRACT TRUNCATED AT 250 WORDS)

Sex and strain differences in the brush border and PAS-positive granules and giant bodies of the mouse renal S3 segment cells.

We recently demonstrated sexual dimorphism in the S3 segment of the ICR mouse kidney, as differences in periodic acid Schiff (PAS) staining on the brush border and the number and size of PAS-positive granules. However, whether these sex dependent features in the S3 segment of the mouse kidney occur only in the ICR strain or are a general feature also observed in other strains is unclear. In the present study, we examined the renal S3 segment of the ICR, BALB/c, C57BL/6, C3H/He and DBA/2 mice strains, which are commonly used in laboratory experiments. PAS staining of the brush border in females of all strains was more intense than that of males, and PAS-positive granules were detected in all females. In male groups, PAS-positive granules were detected in the DBA/2 strain only, but their number was very few. In addition, PAS-positive giant bodies, larger than the nuclear size, were detected in females except those of the C57BL/6 strain. Histometrical investigation demonstrated apparent strain differences in a number of PAS-positive granules and PAS-positive giant bodies. The ultrastructural and cytochemical investigations suggest that the PAS-positive granules and PAS-positive giant bodies were multilamellar lysosomes. We propose that the present findings are significant for comparative morphology in laboratory animal science.

Morphometrical analysis of sex and strain differences in the mouse nephron.

In the present study, we investigated the sexual dimorphisms, effects of castration and sex hormone treatment, and strain differences in the morphology of mouse kidney by measuring the percentage of renal corpuscles with cuboidal cells in the parietal layer of the glomerular capsule, the diameter of renal corpuscles and the number of nuclei of epithelial cells in the proximal convoluted tubule. In ICR mice, the percentage of renal corpuscles with cuboidal cells and the diameter of renal corpuscles were larger in males than in females, and the numbers of nuclei were lower in males than in females. All of these parameters became similar to those of intact females by orchiectomy, and restored to levels similar to those in intact males after testosterone treatment to orchiectomized males. On the other hand, ovariectomy and estradiol treatment gave no effects to any of these parameters. Similar sexual dimorphisms were observed in BALB/c, C57BL/6, C3H/He and DBA/2 mice as in ICR mice, although the diameter of renal corpuscles was not significantly different between males and females in BALB/c mice. Strain differences were observed in each of these three parameters among the 5 strains. In conclusion, the present study demonstrated sexual dimorphisms, effects of castration and sex hormone treatment, and strain differences in the morphology of mouse kidney by morphometrical analysis.

Effects of estrous cycle on the mouse kidney morphology.

Although mice kidney morphology shows various sexual dimorphisms, the effect of the estrous cycle has not previously been discussed. In this study, we investigated the effects of the estrous cycle on kidney morphology, including renin-positive areas, of female DBA/2 mice. No effects were confirmed in most of the histometrical parameters, however, the percentage of the renal corpuscles in which cuboidal epithelium covered under 50% of the parietal layer was significantly higher during estrus compared to that during anestrus.

Staining pattern of the brush border and detection of cytoplasmic granules in the uriniferous tubules of female DBA/2Cr mouse kidney: comparison among various fixations and stains.

The proximal straight tubules of the female mouse kidney exhibit heavy periodic acid Schiff (PAS) staining in their brush borders and numerous cytoplasmic granules. In the present study, the female DBA/2Cr mouse kidney was examined, using various fixatives (formalin, PFA, PLP, Zamboni's, Bouin, or Carnoy solution) and various staining methods (HE, PAS, alcian blue, periodic acid methenamine-silver (PAM), toluidine blue, azan, or Congo red). Under azan and PAM, the staining pattern of the brush border was similar to that of PAS, and few effects of the fixative were observed. Cytoplasmic granules were clearly detected with PAM as well as PAS. However, these granules were not detected with Carnoy solution. Furthermore, distribution of granules differed between PAS and PAM.

Relationship of cerebral blood flow, cerebral metabolism, and electroencephalography to outcome in acute experimental compression ischemia--barbiturate effects on delayed brain swelling.

Acute compression ischemia was induced in 30 cats by progressive inflation of an epidural balloon, followed by rapid decompression. Changes in intracranial pressure, mean arterial blood pressure, cerebral blood flow (CBF), arteriovenous oxygen difference (AVDO2), cerebral metabolic rate of oxygen (CMRO2), and electroencephalography (EEG) were studied in 13 untreated animals and in seven animals receiving barbiturate. Ten cats with cardiovascular or respiratory problems, or intracranial hematoma were excluded from the study. In untreated animals, six (46%) survived with no brain swelling and were classified as the "no swelling group," and seven (56%) died from fatal brain swelling and were classified as the "delayed swelling group." All animals with barbiturate therapy showed no brain swelling. Serial measurement of CBF, AVDO2, and CMRO2 indicated the existence of postischemic delayed hypoperfusion associated with relative hypermetabolism in untreated animals. Good recovery of CBF and CMRO2 was observed in the "no swelling group," and poor recovery in the "delayed swelling group." The time course of the total fast-wave components on EEG was quite similar to that of CMRO2, and the time course of the "CBF index," which is % fast-wave component of EEG divided by AVDO2, was similar to that of CBF. Barbiturates reduced CMRO2 and fast-wave component during administration, possibly improving the relatively hypermetabolic state, and reduced the mortality rate to 0%. The maximum effects of barbiturate could be expected by administering the drug at the stage of delayed hypoperfusion with relative hypermetabolism, indicated by rapid recovery of the % fast-wave component, high AVDO2, and low CBF index.

AVDO2 and EEG as indicators for treatment of post-traumatic brain swelling--experimental study.

A cat model of compression ischemia using epidural balloon inflation investigated: 1) the relationship between postischemic cerebral blood flow (CBF) and metabolism and brain swelling, 2) the use of arteriovenous oxygen difference (AVDO2) and electroencephalographic (EEG) frequency band analysis for monitoring CBF and cerebral metabolism, and 3) indications for selecting the therapy. Global ischemia was induced by brain compression, followed by rapid decompression, and AVDO2, CBF, cerebral metabolic rate for oxygen, and EEG were monitored. The animals were divided into delayed brain swelling and no swelling groups. The severity of compression ischemia influenced the recovery of CBF and cerebral metabolism. The AVDO2 and EEG reflected the CBF and cerebral metabolism. These parameters are useful in selecting the therapy for focal brain injury.

[Experimental and clinical studies of cefmenoxime in the field of obstetrics and gynecology].

The study group was organized to evaluate the usefulness of cefmenoxime (CMX) injection, a new synthetic cephalosporin, for the treatment of infections in the field of obstetrics and gynecology. Fundamental and clinical studies were made by the society and the following results were obtained. 1. The peak distribution of CMX's MIC for E. coli, Klebsiella sp., Enterobacter sp., Bacteroides sp. and Peptococcus sp. isolated from obstetrical and gynecological infections with relatively high frequencies area 0.1, less than or equal to 0.05, 0.2, 3.13, 1.56 micrograms/ml, respectively, with an inoculation of 10(6) cells/ml. 2. When 1 g of CMX is administered by intravenous drip infusion for 1 hour, the maximum concentrations in various tissues of female genital organs were as follows: 14.2 and 13.2 micrograms/g in ovary and oviduct, respectively, at 1.20 hours after the start of administration, and 16.9 and 26.3 micrograms/g in corpus uteri and cervix uteri, respectively, after 1 hour. As for the transfer to the exudate in the pelvic dead cavity, the peak concentration was 15.6 micrograms/ml after 2.13 hours. 3. In the clinical studies, CMX was given to 258 cases with female genital organ infections and others. As for the clinical effects, with exclusion of 3 cases in which other antibiotics are concomitantly used, responses were excellent in 76 cases, good in 162 cases and poor in 17 cases, among 255 cases in total. The efficacy rate was 93.3%. The efficacy rates by diseases were 97.1% (68/70) for intrauterine infections, 88.8% (79/89) for intrapelvic infections, 98.4% (62/63) for adnexitis, and 100% (23/23) for infections of external genital organs. As for the clinical effects on causative bacteria, the efficacy rates were 100% (19/19) for single infections due to Gram-positive bacteria, 94.8% (55/58) for single infections due to Gram-negative bacteria, and 88.2% (15/17) for single infections due to anaerobic bacteria. And its efficacy rates were 89.6% (69/77) for mixed infection cases. Side effects were observed in 2 cases (0.8%); 1 case with eruption, and 1 case with diarrhea and vomiting. As for abnormal laboratory findings, lower white blood cell count was observed in 2 cases and elevation of the values regarding hepatic functions in 9 cases. All cases were returned to the normal after the completion of the administration. Cefmenoxime showed a satisfactory clinical efficacy and a potent bacteriological effect in treatment of the infections in the field of obstetrics and gynecology, and it has been concluded that cefmenoxime will be useful addition to the antibiotics for the therapy of these infections.

[Blood purification after aorto-coronary bypass grafting for dialysis patients].

Four dialysis patients received aorto-coronary bypass grafting (CABG) at Fukuyama cardiovascular hospital from April 1989 to March 1992. We employed continuous ambulatory peritoneal dialysis (CAPD) in two cases, hemodialysis with extracorporeal ultrafiltration method (ECUM + HD) in one case and hemodiafiltration with continuous veno-venous hemofiltration (CVVHF + HDF) in one case for perioperative management. In each cases, intermittent HD was undergone during 4 days before operation. Intraoperatively, HD was used together with extracorporeal circulation. Blood pressure in the postoperative course changed more frequently in CAPD cases than ECUM + HD or CVVHF + HDF case. Especially, CVVHF + HDF stabilized the movement of circulation and kept balance of serum BUN, Cr after the operation (BUN; 27.2 +/- 6.3, Cr; 5.0 +/- 0.6). We concluded that CVVHF + HDF would be the most favorable method to maintain the fluid balance stably for the uremic patients after CABG.

[A case report of BWG syndrome in an elderly patient performed with mitral valve replacement 11 years after single CABG].

A 46-year-old female who had undergone single CABG for BWG syndrome 11 years ago was referred to our hospital for paroxysmal atrial fibrillation. The roentgenogram showed slightly cardiomegaly and the enlargement of LA. Ischemic changes of ECG appeared at I, aVL, V5, V6, as inverted T wave, and low voltage R wave at V1-V4, but non Q wave. The thallium-201 emission computed tomogram at exercise revealed poor perfusion at apical region without redistribution pattern. Catheterization showed mitral regurgitation (grade III), big right coronary artery (RCA) arising from aorta, rich collateral to poor left coronary artery (LCA), and bypass graft was obstructed. The proximal end of LCA was closed, and didn't arise from both pulmonary artery and ascending aorta. In this cases, MVR only without re-CABG to LCA was selected and performed. Postoperative course was uneventful. The result of this case suggested that MVR was an effective surgical procedure for MR of BWG syndrome in the adult case and it was better to add CABG to LCA as much as possible if the ischemic region was large.

[Late malfunction of the Björk-Shiley valve prosthesis due to Delrin disc defacement].

A 55-year-old woman eighteen years after mitral valvular replacement with Delrin disc Björk-Shiley valve prosthesis underwent a reoperation of prosthetic valve replacement for the prosthetic malfunction due to disc defacement. The patient suffered from faintness and vertigo at rest. An echocardiographic examination showed a moderate mitral insufficiency with a normal disc movement. Precise examination on the removed prosthesis revealed accelerated defacement of disc margin which made the ring-disc clearance up to 0.35 mm and strut-shaped groove formation on the inlet surface of the disc occluder. These findings suggested a pronouncedly earlier disc wear than predicted by Björk and co-workers. We concluded, therefore, that a patient undergone a valve replacement with Delrin disc Björk-Shiley valve should be examined periodically by echocardiography even though being without any symptoms.