A comparison between cervical dimensions of pregnant and nonpregnant Santa Gertrudis and Bos taurus cows

Cervical measurements were recorded on palpation per rectum for 603 Bos taurus , Santa Gertrudis, and crossbred cattle. Postmortem cervical measurements were recorded for 77 of these cows. Antemortem measurements were positively correlated with those taken at postmortem examination (cervical diameter: r=0.75, P<0.01 and cervical length: r=0.29, P<0.06). Cervical diameters from antemortem and postmortem specimens were greater in Santa Gertrudis cattle than in Bos taurus and crossbred cattle (P<0.05). Cervical shape was different between Santa Gertrudis cattle and Bos taurus or crossbred cattle (P<0.01), with Santa Gertrudis cows having a higher incidence of conical shaped cervices than other breeds. Within Santa Gertrudis cattle, the incidence of conical cervical shape increased with age (P<0.05). Conical cervical shape was associated with a lower pregnancy rate in all breeds (P<0.01). Within Santa Gertrudis cattle, conical cervical shape was associated with a lower pregnancy rate in 1- to 4-year-old cows (P<0.05) but not in cows more than 4 years of age (P>0.05).     

Mast cell proteases liberate stable encephalitogenic fragments from intact myelin.

Protease-containing supernatants from activated rat mast cells were found to degrade purified rat myelin with a subsequent release of a stable encephalitogenic peptide. The two most abundant peptides were identified as residues 69-87 (GSLPQKSQRTQDENPVV) and residues 69-88 (GSLPQKSQRTQDENPVVH). While additional exposure to the mast cell supernatants removes the COOH terminal histamine from peptide 69-88 to yield peptide 69-87, additional proteolytic degradation of the 69-87 peptide was not detected. Immunization with this peptide emulsified in CFA caused the development of clinical experimental autoimmune encephalomyelitis (EAE) in Lewis rats. In addition this 69-87 sequence was found to activate resting encephalitogenic myelin basic protein-reactive T cell lines to adoptively transfer clinical EAE. The release of stable encephalitogenic peptides from the myelin sheath by mast cell proteases may play a role in activation of encephalitogen-specific T cells during the progression of EAE.     

Role of adherent cells in dextran sulfate-mediated enhancement of mitogen-induced B-cell proliferation

We have studied lipopolysaccharide (LPS)-induced proliferation in the rat and have found that the addition of the compound Dextran sulfate (DxS), which itself is not mitogenic, to LPS stimulated cultures results in significant enhancement of cell division. A "DxS-free" supernatant from DxS stimulated spleen cell cultures is able to substitute for DxS in stimulatory activity. This supernatant possesses interleukin 1 (IL-1) activity, however, the addition of purified recombinant IL-1 to LPS-stimulated cultures does not result in augmentation of proliferation. A DxS-free supernatant from DxS stimulated adherent cells is also able to substitute for DxS in stimulatory activity. The active molecule(s) present in the adherent cell-derived DxS-free culture supernatant appears to be distinct from classical IL-1.     

Release of sphingomyelin phosphodiesterase (acid sphingomyelinase) by ammonium chloride from CL 1D mouse L-cells and human fibroblasts. Partial purification and characterization of the exported enzymes

In cultured human fibroblasts and mouse L-cells the lysosomotropic agent, ammonium chloride, caused release of acid sphingomyelinase into the culture medium. The water-soluble enzymes were partially purified by sequential chromatography on ConA-Sepharose, octyl-Sepharose and Sepharose CL-4B. Mouse sphingomyelinase was purified up to 64-fold and human sphingomyelinase 134-fold from the culture medium. Specific activities were 925 nmol/(h X mg) and 1 434 nmol/(h X mg), respectively. The final enzyme preparations obtained were free of other lysosomal enzyme activities tested and had very similar properties: optimal activity at pH 4.8 (mouse enzyme) and pH 4.4 (human enzyme), Km values of 6.2 X 10(-5)M and 2.4 X 10(-5)M, respectively, and an apparent molecular mass of 68 kDa. In isoelectric focusing the enzymes peaked at pH 4.78 (mouse enzyme) and pH 4.75 (human enzyme).     

Periparturient events in ovariectomized embryo transfer recipient mares

Ovariectomized mares treated with progesterone have established and maintained pregnancy after embryo transfer. This study evaluated the ability of ovariectomized embryo transfer recipients to successfully undergo parturition, raise a foal, and return to a useful reproductive status. Periparturient events in three ovariectomized embryo transfer recipient mares and three intact mares were compared. All mares foaled normally. Mammary scores were similar for both groups and all mares produced sufficient colostrum and milk to allow normal growth of healthy foals. Plasma progesterone levels decreased to < 5 ng/ml by Day 4 post partum in both groups. Progesterone concentrations continued to decrease and remained at <1 ng/ml in ovariectomized mares, but increased after the first postpartum ovulation (Day 9 to 15) in intact mares. Endometrial involution as determined by histological evaluation was complete in ovariectomized mares by Day 10 post partum and in intact mares by Day 11 post partum. As assessed by palpation per rectum and clearance of bacteria from the uterus, uterine involution was similar in all mares. The three ovariectomized mares subsequently received embryos by transcervical transfer and two of them established pregnancy. These results indicate that normal parturition, lactation, maternal behavior and uterine involution are independent of ovarian function.     

The role of mast cells in the elicitation of experimental allergic encephalomyelitis

Experimental allergic encephalomyelitis (EAE), a T cell-mediated autoimmune disease can be transferred with lymphoid cells from actively immunized rats into naive recipients. In the mouse, previous studies have suggested a role for histamine/serotonin in the development of active EAE. We have found that myelin basic protein-reactive cells transfer a biphasic skin test response to naive rats analogous to what has been described in the mouse contact dermatitis system, where mast cell sensitization by Ag-specific T cell factors is required for the induction of skin test responses. Treatment of cell recipients with the serotonin receptor antagonists, cyproheptadine or methysergide, blocked or significantly reduced the development of EAE. Furthermore, it was found that treatment with cyproheptadine was effective in blocking clinical disease when administered day 3 to day 6 after cell transfer. In contrast, cyproheptadine treatments before induction of paralysis day 0 to 3, failed to alter the course of clinical disease. The inhibitor of mast cell degranulation, proxicromil, was also found to effectively block the elicitation of adoptively transferred EAE and was also found to be effective when administered just before the onset of clinical disease. Reserpine, a compound known to deplete mast cells of vasoactive amines by forcing granule contents into the cytoplasm where they are degraded by cell enzymes, was also effective in blocking both active and adoptively transferred EAE. Disease inhibition was found to be partially reversed with pargyline, an inhibitor of monoamine oxidase. In addition lymphocytes from treated animals were capable of transferring disease to naive recipients and appeared to have normal activity as assessed by Ag-or mitogen-driven proliferation in addition to IL-2 production.     

Analysis of reactive carbonyls in the expired air of transgenic mice.

Methods for the determination of trace levels of volatile carbonyl compounds in air expired from mice were developed and validated. Tumor bearing transgenic mice or nontransgenic control mice were placed into a glass chamber through which air was passed continuously at 90 ml/min for 1 h. The effluent gas stream was bubbled into an aqueous cysteamine solution or an aqueous methylhydrazine solution. Formaldehyde, acetaldehyde, and acetone in expired air were derivatized to thiazolidine with cysteamine and malonaldehyde was derivatized to 1-methyl-2-pyrazole with methylhydrazine. The derivatized compounds were analyzed by capillary gas chromatography with flame photometric or nitrogen-phosphorous-specific detection. The lowest level quantitated was 4 micrograms/ml thiazolidine, equivalent to 1.35 micrograms/ml formaldehyde. Formaldehyde was recovered at a level of 1356 +/- 234 nmol/kg0.75 (mean +/- SD) from mice with tumors and 898 +/- 97 nmol/kg0.75 from mice without tumors, suggesting that tumor bearing transgenic mice expired significantly more formaldehyde than did tumor free controls. Amounts of expired acetaldehyde and acetone were not different among mice. Malonaldehyde was not detected in either group of mice.