Examination of ester sulfates in Podzolic and Regosolic soils using an immobilized arylsulfatase reactor

The enzyme kinetics of an immobilized arylsulfatase reactor were examined. We found that the optimum operating conditions for the reactor were pH 7.0 and 25°C, using p-nitrophenyl sulfate in acetate buffer. The Michaelis constant (K _m) of immobilized arylsulfatase was 5.29 mM, compared with a K _m of 2.18 mM for soluble arylsulfatase from the same source (Helix pomatia). Since arylsulfatase hydrolyzes organic ester sulfate linkages, the immobilized arylsulfatase reactor was used to examine ester sulfate compounds in two soils subjected to different fertility management schemes. Soil samples were obtained from the Ap horizons of a Podzol from S-amended wheat plots and a Regosol from dykeland hayfield plots which had received additions of NH₄NO₃ and compost. The distribution of S in these soils was examined in the fall of 1993 and the spring of 1994. Soil organic matter was extracted and separated into three molecular weight fractions (<500, 500–10 000, >10 000). There was no difference in the ester sulfate content for the >10 000 fraction of control and S-amended Podzol soils; however, the S-amended samples had significantly higher quantities of hydrolysable ester sulfates than controls for the 500–10 000 range, indicating that S amendments resulted in the incorporation of ester sulfate into this lower molecular weight fraction. Both control and NH₄NO₃ treatments to the Regosol showed significantly higher quantities of hydrolysable ester sulfates in the >10 000 fraction, while compostamended plots showed no difference between the >10 000 and 500–10 000 fractions due to suspected microbial degradation of high molecular weight organic S compounds in the compost. Since there was no significant effect of sampling time, this study indicated that naturally occurring low molecular weight ester sulfate compounds accumulate in soil and persist during storage. Hydrolysable ester sulfates constituted 35–55% of the hydriodic acid-reducible S in these different soils and probably represent an important and easily mineralizable portion of total ester sulfates.     

Histological changes to the skin of Merino sheep following deep dermal and subcutaneous injections of sodium lauryl sulfate

Objectives To characterise the changes caused to sheep skin by deep dermal and subcutaneous injections of sodium lauryl sulfate (SLS) and describe the subsequent healing process. Procedure On 6 sheep 20 small areas of skin were each given deep dermal and subcutaneous injections of 0.2 mL of 7% SLS. Biopsies were collected at intervals up to 28 days after treatment and the histological changes in each of the treated skin samples were assessed and graded. Results There was no evidence of alterations in behaviour, weight gain or appetite of the sheep following the injections. Initial swelling of the treated site subsided by day 14, leaving a firm, slightly raised crust. At day 21, the treated area was depressed and covered by a scab, which sloughed completely by day 28. There was necrosis of the subcutis and deep dermis 2 min after treatment, followed by inflammation, fibroplasia, angiogenesis and tissue remodelling. Conclusion Injection of SLS caused almost immediate local necrosis followed by eschar formation, sloughing and scarring of treated skin. Deep dermal and subcutaneous SLS is potentially useful as a chemical alternative to mulesing.     

Postanesthetic hemorrhagic myelopathy in a horse

Acute hemorrhagic myelopathy developed in the sixth cervical to the eighth thoracic spinal cord segments of a 1-year-old Quarter Horse colt that was castrated under general anesthesia while in dorsal recumbency. Clinical signs were consistent with severe transverse myelopathy caudal to the brachial enlargement and cranial to the lumbosacral enlargement of the spinal cord. Histologic examination of the spinal cord revealed hemorrhage in the gray matter, with multiple blood-filled clefts in otherwise normal neuropil. Hemodynamic changes in the spinal cord associated with anesthesia and dorsal recumbency may have led to hypoxic vessel damage, with massive hemorrhage after surgery, when the horse was returned to lateral recumbency. Postanesthetic hemorrhagic myelopathy is a possible complication of positioning in dorsal recumbency, during anesthesia, in rapidly growing, young horses.     

Neuromuscular abnormalities associated with hypothyroidism and lymphocytic thyroiditis in three dogs

Various degrees of persistent or paroxysmal paresis involving only the hindlimbs or all four limbs were observed in 3 dogs with hypothyroidism and lymphocytic thyroiditis. Clinical features included lethargy, obesity, alopecia, insidious and progressive paresis, hypotonia, and slow segmental reflexes in 2 dogs. Obesity, alopecia, paroxysmal paresis, and behavior change were observed in the third dog. Laboratory tests indicated that thyroid function was less than normal in all 3 dogs. Abnormal electromyographic potentials and slow motor nerve conduction velocities were found in each dog. Muscle biopsy specimen abnormalities included selective type-II myofiber atrophy in all dogs, whereas one dog had angular atrophy of type-I and type-II myofibers indicative of denervation. A substance that stained with para-aminosalicylic acid was observed within vacuoles of type-I myofibers in one dog. Lymphocytic thyroiditis characterized by lymphocytic infiltration of excised thyroid glands was observed in all dogs.     

Canine mammary mixed tumours: immunohistochemical expressions of EGFR and HER‐2

Background Benign mixed tumours (BMTs) are frequently found in the mammary glands of female dogs, but the factors determining malignant transformation in these tumours are unknown. Objective To evaluate the expression of the oncoproteins, human epidermal growth factor receptor 2 (HER‐2) and epidermal growth factor receptor (EGFR), in 46 carcinomas in BMTs (CBMTs) and to verify their possible association with the malignancy of the tumours. Methods Immunohistochemical expression was analysed in benign and malignant components separately, and then compared with 74 cases of BMTs. Results Among the CBMTs, positivity for HER‐2 was found in the benign histological component of 4.3% (2/46), in the malignant epithelial non‐invasive component of 14.8% (4/27) and in the malignant invasive epithelial component of 13.6% (6/44) of cases. Two of the 24 (8.3%) BMTs were positive for HER‐2. There was no relationship between HER‐2 and the tumour components. There was no significant difference between BMTs and CBMTs. Positivity for EGFR was found in the benign component of 17.4% (8/46) of the CBMTs, in the malignant epithelial non‐invasive component of 40.7% (11/27%) and in the invasive epithelial malignant component of 45.4% (20/44). EGFR positivity was significantly associated with the invasive component of CBMTs. Conclusion EGFR may contribute to malignant epithelial transformation of BMTs. In contrast, HER‐2 overexpression may not be associated with the acquisition of a malignant epithelial phenotype.     

Internal Hydrogen Bond Formation in a syn-Hydroxyepoxide

The existence of an internal hydrogen bond in a compound representative of a syn diol epoxide (a possible intermediate in chemical carcinogenesis by certain polycyclic aromatic hydrocarbons) has been demonstrated by x-ray crystallographic and nuclear magnetic resonance studies. This internal hydrogen bond was found in 3,4-epoxy-2-methyl-1,2,3,4-tetrahydro-1-naphthol and was shown to persist in dioxane-water solutions containing up to 80 mole percent water. In this structure, the 1-hydroxy and 2-methyl groups are shown to occupy axial positions. In the anti diol epoxide, which has no internal hydrogen bond, analogous groups are equatorial. Crystals of the compound were unstable in the x-ray beam while the data were being collected (even at low temperatures), presumably as a result of decomposition.     

Lectin‐binding Sites on the Normal and Pathologic Uterus of Sows

The lectin‐binding pattern was compared in the normal and pathological uterus of sows during the ovarian cycle. The following biotinylated lectins were used: Con A, DBA, SBA, PNA, RCA‐I, UEA‐I and WGA. Glycoconjugate labelling showed differences between phases of ovarian cycle and presence of morphologic lesions. Cystic endometrial hyperplasia increased the RCA‐I reaction in the apical region of the glandular epithelium. There was higher intensity of labelling of WGA in the glandular epithelium in uteri with endometritis. In addition, increased Con A binding in the glandular epithelium and mild reduction of UEA‐I reactivity in the glycocalyx of the glandular epithelium were detected in the cases of endometritis. The results of this study show that morphologic alterations modify the sugar pattern in the porcine uterus. These modifications in glycoconjugates may be one of the reasons for decreased fertility in sows.     

Intermediate Filament Proteins Expression and Carbohydrate Moieties in Trophoblast and Decidual Cells of Mature Cat Placenta

The aim of this study was to characterize cytoskeletal intermediate filament proteins and glycoconjugates of syncytiotrophoblast, cytotrophoblast and decidual cells of feline endotheliochorial placenta. Samples from 12 normal pregnant female cats, after 45 ± 5 days of gestation, were obtained removing the uterine horns by hysterectomy. Sections were processed for routine observation and for immunohistochemistry using anticytokeratin, antivimentin and antidesmin antibodies. In addition, lectin histochemistry was performed using a panel of several biotinylated lectins to characterize glycosides expression profile. Cytotrophoblast and syncytiotrophoblast showed immunoreactivity only with acidic and basic cytokeratins. Decidual cells were only positive to vimentin, consistent with their origin from endometrial fibroblasts. Trophoblast expressed a broad population of glycans, highly exposing terminal N‐acetyl glucosamine residues and non‐sialylated galactose and N‐acetyl galactosamine oligomers. Oligosaccharides bound by Phaseolus vulgaris erythroagglutinin were the only highly branched N‐linked residues evidenced in cats, and they were restricted to the syncytium. Unlike results reported on humans, mice and rats on lectin affinity of decidual cells, sialid acids and complex N‐linked oligosaccharides were not demonstrated in cats. Glycosylation of proteins determines many of their final properties, thus becoming essential for the embryo‐maternal dialogue during implantation and placentation. Changes in glycosylation pattern have been related to pathological pregnancies in other species. Hence, the knowledge about glycosylation profile of the normal cat placenta may lead to a better understanding of both normal and pathological reproductive events.     

Clinical, diagnostic and biochemical features of generalised glycogenosis type II in Brahman cattle

SUMMARY Clinical, diagnostic and biochemical features of generalised glycogenosis are described In 96 Brahman-type calves. Typically the calves were presented when about 6 months of age, with III-thrift and muscular weakness as the most common signs. Acidic α-glucosidase activity was reduced in peripheral blood lymphocytes and skeletal muscle. Muscle glycogen concentration was consistently higher in affected animals than In clinically normal cattle. Other observations in affected calves Included elevation of serum aspartate amlnotransferase and creatine kinase activities and excessive amounts of high molecular weight oligosaccharides in urine. Fine cytoplasmic vacuolation of neurones In the brain and spinal cord, skeletal muscle, myocardlum and of Purkinje fibres were consistent histological observations. Periodic acid-Schlff staining revealed the presence of glycogen-like material In peripheral blood lymphocytes of all affected calves, Indicating that this is a useful aid for the diagnosis of glycogenosis. While 3 of the 96 calves showed somewhat different clinical signs, the similarity of pathology and the biochemical and clinical evidence in the remainder suggested that, In these animals, the disease was expressed as a single syndrome.