The acute toxicity of cadmium to different larval stages of Chironomus riparius (Diptera: Chironomidae) and its ecological significance for pollution regulation

Summary Acute toxicity tests using a static-with-replacement testing procedure were carried out with the four larval instars of the freshwater detritivore Chironomus riparius (Meigen). Median lethal concentrations (10, 24, 48 and 96 h LC50's) indicated great differences in sensitivity to cadmium between instars. Larvae became more tolerant with increasing age, the most resistant stage (fourth instar) having a 24 h LC50 of 2,400 mg Cd l^-1, approximately 950 times greater than the corresponding value of 2.1 mg Cd l^-1 recorded for the most sensitive (first instar) stage. These observations are discussed in relation to the establishment of water quality standards.     

Sequence of a cDNA for mouse thymidylate synthase reveals striking similarity with the prokaryotic enzyme

We report the nucleotide sequence of a cloned cDNA, pMTS-3, that contains a 1-kb insert corresponding to mouse thymidylate synthase (E.C. 2.1.1.45). The open reading frame of 921 nucleotides from the first AUG to the termination codon specifies a protein with a molecular mass of 34,962 daltons. The predicted amino acid sequence is 90% identical with that of the human enzyme. The mouse sequence also has an extremely high degree of similarity (as much as 55% identity) with prokaryotic thymidylate synthase sequences, indicating that thymidylate synthase is among the most highly conserved proteins studied to date. The similarity is especially pronounced (as much as 80% identity) in the 44-amino-acid region encompassing the binding site for deoxyuridylic acid. The cDNA sequence also suggests that mouse thymidylate synthase mRNA lacks a 3' untranslated region, since the termination codon, UAA, is followed immediately by a poly(A) segment.      

The biosynthesis of some androst-16-enes from C21 and C19 steroids in boar testicular and adrenal tissue

1. The formation of androst-16-enes from [4-(14)C]progesterone has been investigated with long-term incubations and short-term kinetic studies. After 4hr., 1.7 and 10.3% respectively of 3alpha- and 3beta-hydroxy-5alpha-androst-16-enes were formed in boar testis minces, but much smaller yields were obtained in boar adrenal. Both tissues formed small quantities of androsta-4,16-dien-3-one. 2. The amounts of androst-4-ene-3,17-dione and testosterone isolated were small, suggesting that androst-16-ene formation may occur preferentially in the boar testis. 3. In the absence of tissue no radioactive androst-16-enes were formed. 4. Incubation of both [4-(14)C]pregnenolone and [7alpha-(3)H]progesterone resulted in 3alpha- and 3beta-hydroxy-5alpha-androst-16-enes containing (3)H/(14)C ratios of near unity and confirmed that both C(21) steroids were precursors. A similar incubation with 17alpha-hydroxy[4-(14)C]-progesterone and [7alpha-(3)H]progesterone gave the same Delta(16)-alcohols, but they contained only (3)H, indicating that side-chain cleavage of pregnenolone and progesterone occurred before 17alpha-hydroxylation. 5. Dehydroepiandrosterone, testosterone, testosterone acetate and 16-dehydroprogesterone were not found to be precursors of Delta(16)-steroids. 6. A pathway is proposed for the biosynthesis of 3alpha- and 3beta-hydroxy-5alpha-androst-16-enes from pregnenolone and progesterone; this may involve androsta-4,16-dien-3-one as an intermediate, but excludes 17alpha-hydroxyprogesterone, testosterone and dehydroepiandrosterone.     

Studies on the biosynthesis in vivo and excretion of 16-unsaturated C19 steroids in the boar

1. In one experiment [7alpha-(3)H]pregnenolone was infused continuously for 12min into the left spermatic artery of a sexually mature boar and blood was collected during this period by continuous drainage from the spermatic vein. After infusion, the testis was removed and immediately cooled to -196 degrees C. 2. From both the testicular tissue and the spermatic venous plasma, (3)H-labelled 16-unsaturated C(19) steroids were isolated and characterized and their radiochemical purity was established. 5alpha-Androst-16-en-3alpha- and 3beta-ol occurred mainly as sulphate conjugates and to a lesser extent as free steroids. Only traces of these alcohols occurred as glucosiduronate conjugates. 5alpha-Androst-16-en-3-one was found in the free (ether-extractable) fraction. 3. The isotope concentration of each of the (3)H-labelled 16-unsaturated C(19) steroids in testicular tissue was different from that in spermatic venous plasma. 4. The ratios of tritiated 5alpha-androst-16-en-3alpha- and 3beta-ol (free steroids) to their respective sulphate conjugates in the testicular tissue were less than the ratios of the same compounds in the spermatic venous plasma. The possibility that the sulphates are partially hydrolysed by testicular sulphatases before secretion is discussed. 5. In a second experiment, a continuous close-arterial infusion of [7alpha-(3)H]pregnenolone into the left testis was performed over a 200min period and all the urine that accumulated during the infusion was collected for analysis. 6. No (3)H-labelled 16-unsaturated C(19) steroids were detected in the urine as free steroids. Only a trace of 5alpha-androst-16-en-3alpha-ol was detected conjugated as glucosiduronate, whereas the corresponding 3beta-alcohol occurred mainly as glucosiduronate and to a lesser extent as sulphate. 7. The absence of 5alpha-androst-16-en-3beta-ol glucosiduronate in the spermatic venous blood and its presence in considerable amount in the urine may be attributed to hepatic glucuronyl transferase activity.     

Studies on the biosynthesis of 16-dehydro steroids: The metabolism of [4-14C]pregnenolone by boar adrenal and testis tissue in vitro

1. The metabolism of [4-(14)C]pregnenolone in vitro by boar adrenocortical and testis tissue has been studied. 2. Boar testis tissue formed three labelled Delta(16)-steroids, 5alpha-androst-16-en-3alpha-ol, 5alpha-androst-16-en-3beta-ol and androsta-4,16-dien-3-one. In adrenal tissue very much smaller yields of the same metabolites were obtained. 3. Both tissues produced labelled progesterone, androst-4-ene-3,17-dione and testosterone in varying quantities. The amount of progesterone was about 120 times greater in the adrenal tissue. In testis tissue dehydroepiandrosterone was found only in small quantity. 4. A pathway is suggested for the biosynthesis of Delta(16)-steroids from pregnenolone in boar testis tissue. The possibility that progesterone may be an intermediate is discussed.     

Aboveground nitrogen and phosphorus use by five plantation-grown trees with different leaf longevities

Aboveground nitrogen (N) and phosphorus (P) requirement, retranslocation and use efficiency were determined for 28-year-old red oak (Quercus rubra L.), European larch (Larix decidua Miller), white pine (Pinus strobes L.), red pine (Pinus resinosa Ait.) and Norway spruce (Picea abies (L) Karst.) plantations on a similar soil in southwestern Wisconsin. Annual aboveground N and P requirements (kg/ha/yr) totaled 126 and 13 for red oak, 86 and 9 for European larch, 80 and 9 for white pine, 38 and 6 for red pine, and 81 and 13 for Norway spruce, respectively. Nitrogen and P retranslocation from current foliage ranged from 81 and 72%, respectively, for European larch, whereas red pine retranslocated the smallest amount of N (13%) and Norway spruce retranslocated the smallest amount of P (18%). In three evergreen species, uptake accounted for 72 to 74% of annual N requirement whereas for two deciduous species retranslocation accounted for 76 to 77% of the annual N requirement. Nitrogen and P use (ANPP/uptake) was more efficient in deciduous species than evergreen species. The results from this common garden experiment demonstrate that differences in N and P cycling among species may result from intrinsic characteristics (e.g. leaf longevity) rather than environmental conditions.     

Inhibition of rat testicular microsomal steroidogenesis by oxytocin and metyrapone

Capillary gas chromatographic 'steroid profiling' has been utilised to separate and quantify the metabolites (derivatized as methyloximes and/or trimethylsilyl ethers) formed from pregnenolone after incubation with rat testicular microsomes. A wide range of steroid metabolites was found, indicating that both the 5-ene and 4-ene pathways of testosterone biosynthesis were operating, as well as 16 alpha-hydroxylation, 20 beta-reduction and the formation of several C19 steroids (the 16-androstenes). At the concentration used, Metyrapone markedly inhibited 16 alpha- and 17-hydroxylation and side-chain cleavage of 17-hydroxylated C21 steroids. 16-Androstene production was also markedly inhibited and the formation of other metabolites was affected to lesser extents. Oxytocin abolished the formation of all C21 and C19 metabolites of pregnenolone.     

Comparative double-blind study of cephalexin and co-trimoxazole in urinary tract infections.

Treatment with cephalexin 1 g twiec daily and cotrimoxazole 2 tablets twice daily was compared in a double-blind, randomised study of 100 women with urinary tract infections. CO-trimoxazole gave a significantly higher cure rate compared with cephalexin two and six weeks after the one-week course of treatment. The higher failure rate with cephalexin was not related to the age of the patient, presentation, pyelographic appearances, or type of organism in the initial infection. Among the failures all but one of the organisms were sensitive to cephalexin. With the dosage regimen and duration of treatment used in this study cotrimoxazole appears to be superior to cephalexin in the management of acute urinary infections.     

Tissue specific O-linked glycosylation of the neural cell adhesion molecule (N-CAM)

We have shown previously that the predominant N-CAM isoform in skeletal muscle myotubes contains as a result of alternative splicing a novel domain (MSD1) in its extracellular region. Here we show that this region represents a site for O-linked carbohydrate attachment. The lipid tailed N-CAM in myotubes was found to bind peanut lectin while the transmembrane isoform from myoblasts lacking MSD1 did not. In addition, N-CAM from a variety of neural sources failed to bind the lectin. Analysis of 3T3 fibroblasts transfected with various N-CAM cDNAs, showed that peanut lectin binding was correlated specifically with the expression of the MSD1 region. The oligosaccharides isolated from a purified preparation of myotube N-CAM were shown to contain an O-linked oligosaccharide whose core structure was a sialylated version of Gal beta 1----3GalNac which is the structure recognized specifically by peanut lectin. These data provide the first evidence for the expression of O-linked carbohydrate on any N-CAM isoform and more specifically target this oligosaccharide to the MSD1 region of myotube N-CAM.