Cloning of a cDNA encoding an Artemia franciscana Na/K ATPase alpha-subunit.

Clones of cDNA that code for an isoform of the Artemia franciscana Na/K ATPase alpha subunit (NaKA alpha) have been isolated. The sequence of the longest of these clones (pArATNa136) is 3595 nucleotides; it codes for a 1004-amino acid protein whose sequence is identical to that of two previously sequenced Artemia NaKA alpha peptides. The encoded protein is over 73% identical to Drosophila melanogaster and vertebrate NaKA alpha s, and 73.8% identical to another Artemia NaKA alpha isoform previously described (named alpha 2850 in this article). The two Artemia cDNA clones code for mRNAs of different size; the clone pArATNa136 codes for a 4.5-kb mRNA while the alpha 2850 clone codes for a 3.6-kb mRNA. The degree of homology and the different size of the mRNAs encoded by both cDNAs suggest that they code for two different isoforms of the protein.     

Effects of progesterone-impregnated sponge treatment on peripheral plasma hormone levels and fertility in the cyclic ewe

Cyclic ewes were treated with 500-mg progesterone-impregnated sponges for the synchronization of oestrus. In the first experiment, the sponges were removed from 79 ewes at intervals over a 17-day treatment and the residual amount of progesterone was measured in order to assess the rate of absorption of the hormone from the sponges by the animals. The residual progesterone was found to decrease linearly with the duration of sponge insertion but there was also a significant quadratic component indicative of a slowing down in the rate of progesterone absorption towards the end of the treatment period. In the second experiment, 13 cyclic ewes were treated with 500-mg progesterone sponges for 17 days and the eight ewes in oestrus following spongewithdrawal were mated. The peripheral plasma progesterone was assayed at intervals during sponge insertion and at weekly intervals after sponge withdrawal. The residual progesterone levels on the sponges and the plasma progesterone levels of the treated ewes were examined in relation to their oestrous response and fertility. There was a significantly higher residual level of hormone remaining on sponges from ewes that mated than on sponges from those that did not (P < 0.01). The 13 ewes exhibited luteal phase levels of plasma progesterone when assayed during the period of sponge insertion regardless of their response to treatment. The mated, fertile ewes had significantly higher plasma hormone levels than the non-mated and the mated infertile ewes, after sponge withdrawal.     

Location of O-acetyl groups in the acidic d-xylan of mimosa scabrella (bracatinga). A study of O-acetyl group migration

Extraction with dimethyl sulfoxide of wood-meal of the stem of bracatinga (Mimosa scabrella), a south Brazilian hardwood, that was defatted and delignified by treatment with aqueous chlorine at 0–5° followed by extraction with cold ethanol, gave a soluble O-acetylated 4-O-methyl-d-glucurono-d-xylan having (1→4)-linked β-d-xylopyranosyl residues that were unsubstituted (65%) and 2-O-(14%), 3-O- (16%), and 2,3-di-O-acetylated (5%), as determined by methylation analysis. Another preparation obtained by use of refluxing ethanol in the delignification process showed neither removal nor migration of acetyl groups. By comparison with synthetic, partly O-acetylated d-xylans of known composition, ¹³C-n.m.r. spectroscopy indicated that O-acetyl group migration does not occur during treatment with cold aqueous chlorine, refluxing ethanol, or water at 70°. Methyl 2-O-acetyl-4-O-methyl-β-d-xylopyranoside (6) was also unaffected by aqueous chlorine. O-Acetyl group migration took place more readily in aqueous and dimethyl sulfoxide solutions of 6 than of O-acetyl-d-xylans. The lowest temperatures at which migration was observed in monosaccharides was at 50 and 70° for solutions in D₂O and (CD₃)₂SO, respectively.     

Four new species of yeast isolated from insect frass in bark ofTsuga heterophylla (Raf.) Sargent

Summary In a study of the yeasts associated with insect frass underneath the bark ofTsuga heterophylla (the Pacific Coast hemlock) four new species of yeast were found. These were described asSporobolomyces singularis, Bullera tsugae, Cryptococcus skinneri andCandida oregonensis. Sporobolomyces singularis is a non-pigmented species, which required an amendment of the genus definition. Ballistospore formation of the new species ofSporobolomyces and ofBullera was absent on malt agar and on potato glucose agar, but positive on corn meal agar. An unusual case of quantitative transgalactosylation by growing cells ofSporobolomyces singularis on lactose has been described. It has been proposed to establish a “Candida parapsilosis Group” of species to whichC. oregonensis was assigned. Supported by a fellowship of the Calouste Gulbenkian Foundation, Lisbon, Portugal.     

The effect of reversion and duplication of leaf disks on the accuracy of photosynthesis determination by the dry weight method

Experiments with sunflower and fodder cabbage showed that reversion of the leaf disks lower side upwards (towards the radiation source) decreased the photosynthetic rate by as much as 26% both at full (2. 10⁵ erg. cm^?2. sec.^?1) and half density of irradiation. On placing two disks one on another the photosynthetic rate was decreased in the upper disk in a similar way as it was on reversion even if it were placed in the normal position (upper side upwards). But assimilation in the lower disk was only a fraction (0–40%) of that of the upper disk and in thick leaves was within the limits of the compensation point. The sum of the increase in dry weight in both disks due to photosynthesis exceeds the increment in one normally placed disk only in thin leaves and at high density of radiation. On the basis of the differences found it is advised to discard experimental samples where a quarter of the disks were reversed during exposure. On evaluating samples in which some of the disks were duplicated in error, it is most accurate to deduct the weight of one control disk from the weight obtained. The decrease in photosynthetic rate found in reversed and duplicated disks is evidently due to the simultaneous effect of differences in the supply of CO₂ and radiation energy to the assimilating tissue and apparatus and also to differences in the photosynthetic capacity of different leaf tissues.     

Investigation of the regulation of pigmentation in alpha-melanocyte-stimulating hormone responsive and unresponsive cultured B16 melanoma cells

In vitro melanocyte-stimulating hormone (MSH) stimulates melanogenesis in some, but not all, melanocytes and melanoma cells. In an attempt to explain this variation in response to alpha MSH, we examined cyclic adenosine monophosphate (cAMP) accumulation, tyrosinase activity, and melanin production in primary (1 degree) murine B16 melanoma cells and in two B16 cell lines (B16 F1 and B16 F10) that are known to respond to alpha MSH. In vivo all three B16 melanoma cell types produced pigmented tumours. In vitro alpha MSH increased tyrosinase activity and melanin content in the F1 and F10 cells but not in the B16 1 degree cells. alpha MSH, however, increased cAMP production in all three cell types, confirming that the inability of B16 1 degree cells to produce melanin in response to alpha MSH is not due to a lack of alpha MSH receptors or cAMP response to alpha MSH. Further, we present evidence for a separate pathway of melanogenesis that is independent of cAMP as calmodulin antagonists, which do not elevate cAMP, increased tyrosinase activity, and melanin production in both 1 degree and F1 cells.     

A new transthyretin mutation associated with amyloid cardiomyopathy.

In transthyretin (TTR) a new mutation (TTR-Thr45) has been identified in a patient with familial amyloidosis characterized clinically by prominent cardiomyopathy and the absence of peripheral neuropathy. Comparative peptide mapping by high-performance liquid chromatography of the patient's plasma TTR together with normal TTR showed the presence of an abnormal tryptic peptide in the patient's TTR. The sequence of this peptide (peptide 6, residues 36-48) demonstrated the presence of a threonine-for-alanine substitution at position 45. This change can be explained by a single base change of adenine for guanine in the Ala-45 codon and was demonstrated directly by DNA sequence analysis of PCR-amplified exon 2 of the TTR gene; allele-specific oligonucleotide hybridization both in the patient and in fixed heart tissue from his aunt confirmed the base change. The TTR-Thr45 mutation is a new variant TTR found associated with cardiomyopathy.