Microbial fermentative preparation of L-[15N2]lysine and its tracer: application to serum amino acid kinetic studies

The microorganism Brevibacterium flavum 21129 has been used to produce multigram batches of L-[15N2]lysine of high purity and isotopic enrichment by supplementation of the growth medium with (15NH4)2SO4 of 98.0 atom% excess. The doubly 15N-labeled lysine can be detected at dilutions 10 times greater than singly labeled lysine when isotope dilution curves are analyzed by gas chromatography-mass spectrometry. This enhanced sensitivity permits kinetic measurements of plasma free-lysine isotope content over a 300-fold dilution during 6 h following a single oral bolus of 5 mg/kg body wt. This inexpensive preparation method lends itself to the production of highly useful biochemical compounds for kinetic studies of human nutrition.     

Muscle development in Caenorhabditis elegans: Mutants exhibiting retarded sarcomere construction

We have studied the structural changes within the body-wall muscle cells of Caenorhabditis elegans during postmitotic development. In wild-type, the number of sarcomeres progressively increases, and each sarcomere appears to grow in length and depth continuously during this period. In mature wild-type cells, the anterior-most body-wall muscle cells have 6–7 sarcomeres; the rest have 9–10 sarcomeres per cell. Twelve mutants in the unc-52 II gene exhibit markedly retarded sarcomere construction and progressive paralysis. Several unc-52 mutants, such as the severely paralyzed SU200, produce only 2–3 sarcomeres per body-wall muscle cell, while the other midly paralyzed unc-52 mutants, such as SU250, build 3–4 sarcomeres per muscle cell. Other structures such as the pharynx and even the noncontractile organelles of the body-wall muscle cells do not appear to be structurally or functionally altered. The unc-52 body-wall sarcomeres become moderately disorganized as they are outstripped by cell growth; sufficient order is preserved, however, so that the majority of thick and thin filaments still interdigitate.The myosin heavy chains of SU200 body-wall muscle fail to accumulate normally, while the pharyngeal myosin heavy chains do not appear to be specifically affected. This biochemical result correlates well with the specificity of morphological changes in the mutant. A model is discussed in which the biochemical and morphological deficits are explained by a simple regulatory mechanism.     

Plasma relaxin concentrations in the pig during the periparturient period: Association with prolactin,estrogen and progesterone concentrations

Concentrations of relaxin, prolactin, unchromatographed estradiol 17_β (E_2β) and progesterone (P₄) were measured in serial samples of inferior vena caval blood, in three pigs, during late pregnancy, and parturition. Maximal relaxin concentrations occurred 60 to 24h before parturition, and ranged from 60 to 286ng/ml. Prolactin concentrations increased from 12.5ng/ml, 48 to 36 hours before parturition, to between 79 to 184ng/ml. At the time of the relaxin surge, E_2β levels were high, and P₄ concentrations were decreasing, thus raising the $\text{E}{}_{\mathrm{2\beta }}\text{P}{}_4$ ratio. A surge in prolactin concentrations followed upon a decline of P₄ to less than 10ng/ml, coinciding with the increase in relaxin concentrations in 2 gilts, and following the surge in relaxin in the third. Udder development occurred near the time of increased relaxin concentrations. ‘Milk let down’ followed maximal relaxin and prolactin concentrations in two gilts, and the increase in prolactin, rather than the increase in relaxin concentration, in the third.     

Successful long-term cryopreservation of neonatal rat islet tissue

Neonatal rat pancreatic tissue was frozen to -196 degrees C using Me2SO as a cryoprotectant and a slow freezing rate to -70 degrees C followed by immersion in liquid nitrogen. Rapid thawing was used. Viability was demonstrated by successful transplantation to streptozotocin-induced diabetic recipients. Long-term preservation, up to 4 weeks, did not demonstrably affect viability. Cryopreservation techniques may afford a method for providing a diabetic recipient the opportunity to receive a large quantity of pooled islet tissue from well-matched donors.