Polymerase chain reaction analysis of mitochondrial DNA polymorphism in N'Dama and Zebu cattle

A study of polymorphisms of mitochondrial DNA (mtDNA) of West African N'Dama (Bos taurus) and East African Zebu (B. indicus) cattle was carried out to obtain information on maternal phylogenetic relationships between these breeds. A relatively large sample size was made possible by using polymerase chain reaction (PCR) amplification of DNA prepared from small blood samples to generate fragments of two known polymorphic mtDNA regions, one within the gene encoding subunit 5 of NADH dehydrogenase and one encompassing the entire D-loop. This approach allowed us to achieve a higher resolution restriction analysis on mtDNA from more animals than would have been possible by conventional methods. PCR-amplified mtDNA of 58 animals from five populations was examined at 26 restriction sites by 16 enzymes. In this way 154 nucleotides of mtDNA were scanned for polymorphism. Six polymorphic sites were located by this means, five of which were within the D-loop and one of which was within the NADH dehydrogenase 5 gene. None of the polymorphisms observed could be con sidered typical of breed or type.     

4-Thiaproline reduces heart lipid peroxidation and collagen accumulation in the diabetic db/db mouse

Summary Collagen accumulation is a main pathological feature of diabetic cardiomyopathy. The underlying mechanisms seem to be increased cross linking by reactive carbonyles. The purpose of the study was to decrease the collagen content of total ventricular tissue by the oral administration of thiaproline, which could reduce collagen due to its functions as a proline analogue, blocking collagen production and as a free oxygen radical scavenger, blocking reactive carbonyles and oxygen species and subsequently collagen cross linking.Thiaproline was administered to genetically diabetic db/db mice and compared to untreated animals. Total ventricular collagen as expressed by hydroxyproline was significantly lower in the treated group (means 0.23 micromoles/10 tissue in the treated vs 0.35 micromoles/100 mg tissue in the untreated group, p < 0.001). Significantly more collagen could be eluted in the treated group (p < 0.001) and carboxymethyllysine was significantly reduced in the treated group (p < 0.001). Di-tyrosine and glycemic control did not differ between the groups. Glutathione was significantly increased in the TP treated experimental group (p < 0.001) and lipid peroxidation products were significantly decreased (means 0.221 absorbance in the treated group versus 0.321 absorbance in the untreated diabetic group) correlating with total ventricular collagen content (r = 0.87, p < 0.01).We conclude that thiaproline reduced total ventricular collagen content by inhibiting collagen cross linking as reflected by increased solubility of collagen and expressed by higher elution quantity of collagen. Thiaproline, and/or its metabolites induced increase of heart glutathione which may well have been scavenging reactive carbonyles derived from lipid peroxidation and advanced stage nonenzymatic glycosylation as shown by decreased total ventricular carboxy-methyllysine and lipid peroxidation products paralleling reduced heart collagen content.It remains to be shown that the successful reduction of heart collagen by thiaproline is paralleled by improved functional properties.     

The expression of Plasmodium falciparum bloodstage antigens in Escherichia coli

A library of cDNA clones expressing proteins of the asexual blood stages of a Papua New Guinean isolate of Plasmodium falciparum (isolate FCQ27/PNG (FC27] was constructed in the bacteriophage vector lambda gt11-Amp3. In an in situ colony immunoassay, human serum was used to identify colonies producing natural immunogens. Sera from donors of defined clinical status, or reactive to a defined subset of natural immunogens were used to identify clones of particular interest (for example, clones reacting with convalescent but not with acute serum or clones expressing the isolate specific S-antigen of FC27). Antisera raised by immunizing mice and rabbits with cloned antigens were used to characterize the P. falciparum proteins corresponding to the antigen-positive clones. Nucleotide sequence analysis of an antigen found on the surface of cells infected with ring stage parasites revealed an unusual sequence coding for eight, four and three amino acid repeats rich in acidic amino acids. The discussion centres on the use of cloned antigens as tools for the analysis of the host-protective immune response and selection of candidate vaccine molecules.     

The duration of ovulation in pigs, studied by transrectal ultrasonography, is not related to early embryonic diversity

The duration of ovulation in pigs was studied by transrectal ultrasonography. The number of preovulatory follicles was counted on both ovaries at 30-minute intervals from 36 hours after the onset of estrus (Group A: naturally ovulating sows that were group-housed and were inseminated and caged during scanning) or 40 hours after treatment with human chorionic gonadotropin (hCG) (Group B: tethered sows that had been induced to ovulate but were not inseminated). The duration of ovulation was (mean+/-SD) 1.8+/-0.6 hours (range 0.75 to 3.25) in Group A (n=13) and 4.6+/-1.7 hours (range 2.0 to 7.0) in Group B (n=8). The difference was significant (P<0.01). In Group A and B sows, respectively, the course of ovulation, expressed as the relation between the relative follicle count (percentage of the maximum follicle count; Y) and the time (percentage of the duration of ovulation; X) was: Y = 104.3( *)e(-0.023( *)X) (R(2)=0.95) and Y = 98.9( *)e(-0.018( *)X) (R(2)=0.92). The onset of ovulation occurred at approximately two-thirds of the duration of the estrus (Group A: 67+/-6%; Group B: 60+/-10%). Group A sows were artificially inseminated and were slaughtered at 98+/-8 hours (range 77 to 110) after ovulation. The difference between the maximum follicle count and the corpora lutea count was zero or only 1 in 81% (21 26 ) of the ovaries. Embryonic diversity (within-litter SD of the number of nuclei or of the number of cell cycles) was not related to the duration of ovulation, neither at the level of ovary nor of sow (P>0.05). In conclusion, transrectal ultrasonography was found to be an appropriate nonsurgical method of studying the duration of ovulation in pigs. The duration of ovulation varied both between sows and between groups of sows, and was not related to early embryonic diversity.     

External and internal factors regulating metabolic rates of an estuarine benthic community

Summary The structure and metabolism of a soft-sediment estuarine macrofaunal community were measured over an annual cycle at two depth-contours in mesohaline Chesapeake Bay. Additional data for plankton productivity and respiration, as well as seston and sediment organics are also summarized for these communities. Benthic community respiration ranged from 0.24–3.38 g O₂ m^-2 d^-1, and significant differences were detected between the two depths. Similarly, macroinfaunal standing stocks reached 11.2 and 32.3 g (ash free) m^-2 for 3 m and 6 m depth communities, respectively, and both exhibited mid-summer declines in abundance. Inferences drawn from these data facilitated a partitioning of benthic community respiration into macrofaunal and meiofaunal/microbial components with a residual term, much of which could be explained statistically by interactions between these two components. A multi-variate statistical model developed from these data matched benthic respiration measurements within 1–2 S.E. Mass-balances of organic carbon were estimated for water column and benthos at the two depthcontours for early and late summer, as well as for an entire, time-weighted year. These various analyses led to the tentative conclusions that this benthic community was regulated by such internal factors as macrofaunal/meiofaunal grazing and microbial gardening, and by external factors such as temperature and predation by nekton. However, it appears that the ultimate control for this community was the supply of energy from organic carbon.Contribution No. HPEL-1206, USASupported by grants with the Maryland Department of Natural Resources (PS-72-02(77-78)), J.A. Mihursky, Coordinating Principal Investigator     

Aspects of the structure and functional anatomy of the Middle Triassic cynodont Luangwa

The study concerns an incomplete skull and postcranial skeleton of the traversodontid cynodont Luangwa drysdalli Brink, from the Middle Triassic Ntawere Formation of Zambia. The incisor teeth had cutting edges maintained by tooth abrasion, and the post-canines functioned more by a puncture-crushing mechanism than by shearing. The jaw muscles, in conjunction with the teeth, were arranged to avoid stresses at the jaw articulation.
Both the region of the orbit, and the braincase show certain tendencies towards mammalian structure.
The forelimb operated in a primitive, sprawling fashion. The equivalent of the therian supraspinatus muscle had evolved, but not the infraspinatus. The hindlimb was more or less erect, and the hip musculature was very close to mammalian. Expanded costal plates were still present on the posterior thoracic and lumbar ribs, and functioned to maintain the vertebral column virtually rigid.