Sex differences in nutritional modulation of gonadotropin secretion during development: studies in the growth-retarded lamb

This study determined whether changes in nutrition during development alter LH secretion in males in a manner similar to that in females; sheep were used as an experimental model. Studies were conducted in the absence of gonadal steroid negative feedback. First, we compared the effect of chronic growth restriction on LH secretion in male and female lambs. Second, we determined whether the gonadotropic response to acute increases and decreases in nutrition is sexually differentiated. Seven male and 8 female Suffolk lambs, gonadectomized, and weaned by 8 wk of age were maintained at a target weight of 20 kg by level of nutrition. After 7 wk of chronic low nutrition (15 wk of age), LH pulse frequency was equally low in males (2.0 +/- 0.7 pulses/4 h) and females (2.0 +/- 0.4 pulses/4 h) relative to that (ca. hourly pulses) in normally growing gonadectomized lambs. Seven weeks later, at 22 wk of age, LH pulse frequency dropped further (males 0.9 +/- 0.3/4 h; females 0.9 +/- 0.4 pulses/4 h). The results of this first experiment, in which we observed no sex difference in gonadotropin secretion under chronic growth restriction, imply equal neuroendocrine sensitivity in males and females to long-term low nutrition. In the second experiment, however, a sex difference was evident in the response to increased and decreased nutrition. Both sexes responded to feeding ad libitum with a rapid increase in LH pulse frequency, but the response was greater in the males than in the females.(ABSTRACT TRUNCATED AT 250 WORDS)     

Screening of insect cell lines for the production of recombinant proteins and infectious virus in the baculovirus expression system.

Eight cell lines derived from the insects Spodoptera frugiperda, Trichoplusia ni, Mamestra brassicae, and Estigmene acrea were evaluated for recombinant beta-galactosidase and infectious virus production following infection with the baculovirus Autographa californica multiple nuclear polyhedrosis virus (AcMNPV). Production was assessed on a specific (per cell and per microgram of uninfected cellular protein) and on a volumetric (per milliliter) basis. Cell density was found to be an important factor in comparing the cell lines due to a density-dependent inhibition of specific protein and virus production that appeared to result from cell-cell contact. After infection of cells at low-density specific beta-galactosidase production per cell would drop between 3- and 6-fold in five of the eight cell lines when plated on tissue culture plates at near-confluent and confluent cell densities. The cell lines Sf 21 and Sf 9 were least sensitive to cell density. After accounting for cell density effects and differences in cell size, two cell lines, BTI Tn 5B1-4 and BTI TnM, were identified that were superior to the other cell lines, including Sf 21 and Sf 9, in beta-galactosidase production. Optimal volumetric and specific beta-galactosidase production from Tn 5B1-4 and TnM cells was 2-fold and 5-fold higher, respectively, in both cell lines than the optimal production from Sf 9 or Sf 21 cells. The Tn 5B1-4 cell line also had the highest viability of all the cell lines at 3 days postinfection and could be adapted to serum-free media.(ABSTRACT TRUNCATED AT 250 WORDS)     

Preparation and properties of recombinant DNA derived tobacco mosaic virus coat protein

Recombinant DNA derived tobacco mosaic virus (vulgare strain) coat protein (r-TMVP) was obtained by cloning and expression in Escherichia coli and was purified by column chromatography, self-assembly polymerization, and precipitation. SDS-PAGE, amino terminal sequencing, and immunoblotting with polyclonal antibodies raised against TMVP confirmed the identify and purity of the recombinant protein. Isoelectric focusing in 8 M urea and fast atom bombardment mass spectrometry demonstrated that the r-TMVP is not acetylated at the amino terminus, unlike the wild-type protein isolated from the tobacco plant derived virus. The characterization of r-TMVP with regard to its self-assembly properties revealed reversible endothermic polymerization as studied by analytical ultracentrifugation, circular dichroism, and electron microscopy. However, the details of the assembly process differed from those of the wild-type protein. At neutral pH, low ionic strength, and 20 degrees C, TMVP forms a 20S two-turn helical rod that acts as a nucleus for further assembly with RNA and additional TMVP to form TMV. Under more acidic conditions, this 20S structure also acts as a nucleus for protein self-assembly to form viruslike RNA-free rods. The r-TMVP that is not acetylated carries an extra positive charge at the amino terminus and does not appear to form the 20S nucleus. Instead, it forms a 28S four-layer structure, which resembles in size and structure the dimer of the bilayer disk formed by the wild-type protein at pH 8.0, high ionic strength, and 20 degrees C.(ABSTRACT TRUNCATED AT 250 WORDS)     

Mechanistic studies of ionizing radiation and oxidative mutagenesis: genetic effects of a single 8-hydroxyguanine (7-hydro-8-oxoguanine) residue inserted at a unique site in a viral genome

T4 RNA ligase was used to construct a deoxypentanucleotide containing a single 8-hydroxyguanine (7-hydro-8-oxoguanine; G8-OH) residue, which is one of the putatively mutagenic DNA adducts produced by oxidants and ionizing radiation. The pentamer d(GCTAG8-OH)p was prepared by the ligation of a chemically synthesized acceptor molecule, d(GCTA), to an adducted donor, 8-hydroxy-2'-deoxyguanosine 5',3'-bisphosphate. The acceptor was efficiently converted to the reaction product (greater than 95%), and the final product yield was 50%. Following 3'-dephosphorylation, the pentamer was characterized by UV spectroscopy, by high-pressure liquid chromatography, and by gas chromatography-mass spectrometry of the nucleosides released by enzymatic hydrolysis. Both d(GCTAG8-OH) and an unmodified control were 5'-phosphorylated by using [gamma -32P]ATP and incorporated covalently by DNA ligase into a five-base gap at a unique NheI restriction site in the otherwise duplex genome of an M13mp19 derivative. The ligation product contained G8-OH at the 3' residue of an in-frame amber codon (5'-TAG-3') (genome position 6276) of the phage lacZ alpha gene. The adduct was part of a nonsense codon in a unique restriction site in order to facilitate the identification and selection of mutants generated by the replication of the modified genome in Escherichia coli. Both control and adducted pentamers ligated into the genome at 50% of the maximum theoretical efficiency, and nearly all (approximately 90%) of the site-specifically adducted products possessed pentanucleotides that were covalently linked at both 5' and 3' termini. The G8-OH lesion in the NheI site inhibited the cleavage of the site by a 200-fold excess of NheI.(ABSTRACT TRUNCATED AT 250 WORDS)     

Prevention of thromboxane B2-induced hepatocyte plasma membrane bleb formation by certain prostaglandins and a proteinase inhibitor

Isolated hepatocytes incubated in the presence of thromboxane B2 developed many plasma membrane blebs which are a characteristic feature of toxic or ischaemic cell injury. When hepatocytes were incubated in the presence of both thromboxane B2 and the non-lysosomal proteinase inhibitor, leupeptin, were also well protected from the formation of blebs. This implies that thromboxane B2 is able to activate non-lysosomal proteinases which appear to attack certain cytoskeletal proteins. The data presented are consistent with thromboxane B2 acting as an intermediary in a proposed mechanism of cell injury and death in which elevated cytosolic free Ca2+ levels activate phospholipase A2 and the arachidonic acid cascade.     

Birth spacing patterns in humans and apes

Comparative studies of birth interval dynamics in wild primates suffer from several problems of analysis and interpretation: (1) the data are always right-censored, (2) sample sizes are usually small, (3) the distribution of birth intervals is expected to be non-normal, (4) early offspring mortality is a confounding variable, and (5) differences in life history (e.g., presence or absence of menopause) can complicate interpretation of the results. A survival analysis designed to minimize these problems is applied to published data on wild chimpanzees and gorillas from Gombe and Virunga Parks, respectively, and to new data on wild orangutans from Tanjung Puting National Park and on a human population, the Gainj of highland Papua New Guinea. According to this analysis, the estimated median birth interval (when the offspring whose birth opens the interval does not die within the interval) is 43.3 +/- 1.0 months for the Gainj, 45.5 +/- 1.2 months for gorillas, 66.6 +/- 1.3 months for chimpanzees, and 92.6 +/- 2.4 months for orangutans.     

Metrical analysis of the basicranium of extant hominoids and Australopithecus

The size and shape of the basicranium (seen in norma basilaris) in Homo, Gorilla, Pan, Pongo, and Australopithecus have been studied by recording the relative disposition of midline and bilateral bony landmarks. Fifteen linear measurements and two angles were used to relate the landmarks. The relatively longer and narrower cranial base of Gorilla, Pan, and Pongo is clearly contrasted with the wider, shorter cranial base in Homo sapiens. When the same observations were made on two “robust” and two “gracile” australopithecine crania, marked differences were found between the taxa. In the two “robust” specimens, the foramen magnum is located relatively further forward, and the axis of the petrous temporal bone is aligned more nearly with the coronal plane than in the two “gracile” crania. The implications of this apparent parallelism in basicranial morphology between Homo sapiens and the “robust” australopithecines are discussed.     

Location of a gene conferring resistance to knockdown by permethrin and bioresmethrin in adults of the BKPM3 strain of Aedes aegypti

A genetic analysis of resistance to knockdown by permethrin and bioresmethrin in adults of the homozygous resistant strain of Aedes aegypti, BKPM3, was made. A major resistance gene R ^pywas located approximately 13 cross-over units from the linkage group III marker, blt (black tarsus).     

Non-Hodgkin's lymphoma: identification of the monoclonal B lymphocyte component in the presence of polyclonal immunoglobulin

A high proportion of non-Hodgkin's lymphomas are neoplastic proliferations of B lymphocytes, and, as such, express integral membrane and/or cytoplasmic immunoglobulin (Ig). Because these cellular proliferations are monoclonal, the Ig of all neoplastic lymphocytes will have identical light chain type and idiotype. These tumors sometimes contain significant amounts of polyclonal Ig. In this study we demonstrate that the polyclonal non-B-lymphocyte-associated Ig may be removed by washing tissue at low pH to reveal either neoplastic B lymphocytes or neoplastic "null" lymphocytes. These observations should facilitate the application of immunohistology to the routine diagnosis of lymphoma.