Blood oxygen transport and hemoglobin function in three tropical fish species from northern Australian freshwater billabongs

The oxygen binding properties of whole blood and hemoglobin were studied in three phylogenetically distant tropical teleosts inhabiting freshwater billabongs that exhibit marked seasonal oxygen stratification. The water breathing saratoga (Scleropages jardinii, Family Osteoglossidae), derived from an ancient divergence of the stem line leading to all other extant teleosts, has a low blood oxygen carrying capacity, Bohr and Root effect, and a high blood oxygen affinity. Erythrocytes contain a single major hemoglobin component, and equimolar concentrations of ATP and GTP. The tarpon (Megalops cyprinoides, Family Megalopidae), a facultative air-breather, has the highest blood oxygen carrying capacity and Bohr effect, and a low blood oxygen affinity and Root effect. Erythrocytes contain a single major hemoglobin and ATP as a putative regulator of hemoglobin oxygen affinity. Barramundi (Lates calcarifer, Family Centropomidae), is an obligate water-breather with intermediate blood oxygen binding properties, and the smallest Root effect. Erythrocytes contain at least 7 hemoglobins, and equimolar concentrations of ATP and GTP. Functional properties of these three blood oxygen transport systems are considered in terms of the respiratory environment and demand for oxygen. Our interpretation supports the hypothesis that the process of speciation can lead to divergence in physiological mechanisms, irrespective of past or present selection pressures.     

Fundamentals of host immune response against Brucella abortus: what the mouse model has revealed about control of infection

The studies reviewed here evaluated the role cellular immune system components play in control of brucellosis by conducting comparative studies with brucella-resistant C57BL/10 or C57BL/6 mice and susceptible BALB/c mice. We have shown by both in vitro and in vivo studies that activation of macrophages with interferon-gamma (IFN-γ) is an important factor for control of infection with B. abortus in the mouse model and that the mechanism of anti-brucella activity largely involved reactive oxygen intermediates. Differences in control of the organism by resistant and susceptible mice was not related to inherent differences in the ability of their macrophages to control infection either with or without IFN-γ activation nor was it attributable to NK cells since we found no role for them in control of brucellosis in either mouse strain. However, relative resistance to brucellosis did correlate with increased production of IFN-γ by CD4 T cells during the first weeks after infection while IL-10 contributed to susceptibility in BALB/c mice. Moreover, by 3 weeks post-infection splenocytes from the susceptible BALB/c mice failed to produce IFN-γ and relied on TNF- as well as CD8 T cells to control infection until the end of the plateau phase around 6 weeks post-infection when IFN-γ production resumed and clearance began. In contrast, IFN-γ was crucial for control throughout the infection in the more resistant C57BL/6 mice and the mice died in its absence by 6 weeks post-infection compared to 12 weeks for the more susceptible mice that relied on additional mechanisms of control. In contrast to the IFN-γ knock-out mice, both β2 microglobulin knock-out C57BL/6 mice, which do not express conventional MHC class I molecules and thus cannot present antigen to CD8 T cells, or perforin knock-out C57BL/6 mice, which have no T cell cytotoxic activity, controlled and cleared the infection as well as normal C57BL/6 mice. The hiatus of IFN-γ production in BALB/c mice correlated with very high levels of total IL-12 and it was postulated that the lack of IFN-γ was a consequence of p40 homodimer blocking activity. However, reduction of p40 IL-12 in vivo through administration of indomethacin reduced the infection without a concomitant measurable increase in IFN-γ. Current studies are aimed at elucidating the mechanism of the IFN-γ hiatus.     

Peritoneal fluid accumulation in the femoral canal as a result of a post-traumatic femoral hydrocele in a mature Warmblood stallion

The following case report documents the clinical presentation, ultrasonographic appearance, surgical management and outcome of a horse with a traumatic tear extending from the superficial and deep inguinal rings, across the aponeurosis of the external abdominal oblique muscle and inguinal ligament, disrupting the femoral ring, vascular lacuna and peritoneum. This subsequently resulted in fluid accumulation within the femoral canal. This unusual injury and presentation gives rise to discussion of the aetiology and nomenclature that should be used to properly describe this injury.     

Technical note: isolation and characterization of sheep ruminal epithelial cells

A system for the isolation and characterization of sheep ruminal epithelial cells has been developed. Ruminal papillae from the ventral cranial sac of 12- to 24-wk-old Dorset ram lambs were subjected to serial tryptic digestions. Initial digestions (two 15-min periods) in the series were used to remove keratinized cells of the stratum corneum, which were able to produce only small amounts of beta-hydroxybutyrate from butyrate (5.37 +/- .72 nmol/120 min/per milligram of dry cell weight). Later digest fractions, containing primarily cells from the stratum basale, exhibited high viabilities (75 to 95%) and proved capable of converting butyrate to beta-hydroxybutyrate at relatively high rates (33.6 +/- 6.7 nmol/120 min per milligram of dry cell weight). Neither acetate nor propionate underwent significant conversion to beta-hydroxybutyrate. However, addition of acetate inhibited (77.4% of control) and addition of propionate stimulated (200% of control) beta-hydroxybutyrate production. Acetate addition reduced the propionate-induced stimulation of beta-hydroxybutyrate production from butyrate (136% of control). These results are similar to those obtained from in vitro incubations of ruminal papillae and suggest that an isolated cell system may prove useful in the further study of ruminal epithelial metabolism.     

Seroprevalence and comparison of isolates of endometriotropic bovine herpesvirus-4.

Sixty-eight cases of suppurative, ulcerative endometritis associated with Bovine Herpesvirus-4 (BHV-4) in postparturient dairy cows (62 Holsteins and 6 Jerseys, mean age 4.2 years) were confirmed by a combination of histopathology, fluorescent antibody assays, electron microscopic evaluation of uterus, and polymerase chain reaction (PCR). All cases occurred in the 3- to 28-day postpartum period, and histologic lesions among various cows were consistent when compared with postpartum interval. The endometrial lining epithelium was necrotic and ulcerated from 3 to 7 days postpartum, with only mild inflammation in the lamina propria and submucosa. From 1 to 4 weeks postpartum, the ulcers were confluent to diffuse. Epithelium was replaced by fibrinonecrotic, suppurative mats, resulting in severe bacterial pyometra by day 24. Seroprevalence to BHV-4 in one dairy with a history of 18 mortality cases was 36% (107 of 296). In a random sample of 8 cows from this herd, none had serologic titers in blood sampled 2 weeks prepartum, but 3 of 8 seroconverted with significant titers of 1:8 to 1:16 at 2 weeks postpartum. By 10 weeks postpartum, all 8 cows returned to negative serologic status. Two of 6 cats from the premises also had positive titers. Random serum samples taken from 480 dairy cattle at sale barns indicated 76 (16%) were positive by serum neutralization. Clinical signs, postparturient timing, and histologic lesions were very similar to those previously reported in Belgium with BHV-4. But sequence analysis of PCR products of the glycoprotein B region of 4 separate field isolates of endometriotropic BHV-4 suggests these field isolates were more closely related to the North American nonvirulent strain DN-599 than to the endometriotropic European strain V.     

Animal performance, plasma hormones and metabolites in Holstein and Belgian Blue growing-fattening bulls

Six Holstein (light-muscled type) and six Belgian Blue bulls (double-muscled type) were fed a finishing diet. Average daily gain was 1.36 kg for the Holstein bulls vs 1.24 kg for the Belgian Blue bulls (P less than .05). Holstein bulls consumed more feed (2.3 vs 1.8 kg/100 kg body weight, P less than .001) than the Belgian Blue bulls. The dressing percentage (55.4 vs 65.8%, P less than .001) and the proportion of muscle (56.1 vs 71.3%, P less than .001) in the carcass were less, whereas the proportions of adipose tissue (28.3 vs 15.4%, P less than .001) and bone (15.7 vs 13.4%, P less than .05) were higher in the Holstein bulls. Plasma creatinine determined in samples obtained once a week was lower (11.0 vs 20.3 mg/liter, P less than .001) in the Holstein bulls. In contrast, Holstein bulls tended to produce more triiodothyronine (2.3 vs 1.8 nM, P less than .10), tetraiodothyronine (71.9 vs 54.7 nM, P less than .10) and insulin-like growth factor I (IGF-I; 340 vs 205 ng/ml, P less than .20) than the Belgian Blue bulls. Growth hormone, insulin, IGF-I and testosterone were measured at 20-min intervals during two 24-h periods. In wk 6, Holstein bulls tended to produce more growth hormone than the Belgian Blues, as indicated by higher total peak area (3,185 vs 2,431 ng), peak amplitude (34.1 vs 22.6 ng/ml, P less than .10) and baseline (4.6 vs 3.3 ng/ml, P less than .20). In wk 27, the trends were opposite.(ABSTRACT TRUNCATED AT 250 WORDS)     

Rapid HPLC screening of jasmonate-induced increases in tobacco alkaloids, phenolics, and diterpene glycosides in Nicotiana attenuata

A rapid, HPLC-based screening procedure for the main classes of secondary metabolites in Nicotiana attenuata leaves (alkaloids, phenolics, and diterpene glycosides) is reported. In a single step, leaves are extracted in aqueous acidified (0.5% acetic acid) methanol, and the extracted compounds are separated by reversed-phase HPLC with an acidic water/acetonitrile gradient in <30 min. The utility of the method in quantifying changes in the secondary metabolites after methyl jasmonate treatment of the plants, a treatment known to elicit resistance to herbivores in nature, is illustrated. Methyl jasmonate treatment elicited dramatic increases in some secondary metabolites (caffeoylputrescine, nicotine, and diterpene glycosides increased 12.5-, 1.4-, and 1.9-fold, respectively) but left others, such as rutin, unchanged. Such broad-based analytical screens will help characterize environmental and genetic changes in secondary metabolite profiles.     

Heterogeneous atmospheric reactions: sulfuric Acid aerosols as tropospheric sinks

The collisional reaction probabilities of several atmospheric species on bulk sulfuric acid surfaces indicate that heterogeneous processes may be important in tropospheric chemistry.     

Conserved folding in retroviral proteases: crystal structure of a synthetic HIV-1 protease

The rational design of drugs that can inhibit the action of viral proteases depends on obtaining accurate structures of these enzymes. The crystal structure of chemically synthesized HIV-1 protease has been determined at 2.8 angstrom resolution (R factor of 0.184) with the use of a model based on the Rous sarcoma virus protease structure. In this enzymatically active protein, the cysteines were replaced by alpha-amino-n-butyric acid, a nongenetically coded amino acid. This structure, in which all 99 amino acids were located, differs in several important details from that reported previously by others. The interface between the identical subunits forming the active protease dimer is composed of four well-ordered beta strands from both the amino and carboxyl termini and residues 86 to 94 have a helical conformation. The observed arrangement of the dimer interface suggests possible designs for dimerization inhibitors.