Species identity of insect cell lines

Summary Three mosquito cell cultures designated as Suitor's clone ofAedes aegypti, Culiseta inornata andAedes vexans were shown to be moth by immunological, karyological, and isozyme analyses. The cells reacted with rabbit antimoth serum but not rabbit antimosquito serum. Chromosome analyses indicated Lepidopteran rather than Dipteran morphology, and three isozyme systems were confirmative. Any one of these assays would be sufficient to indicate that contamination had occurred and could be used as a periodic check for identity of cell cultures. Morphology and growth characteristics are also valid criteria to distinguish between these particular orders of insect cells. These studies were supported by Grant CA-04953-12 from the National Cancer Institute; General Research Support Grant FR-5582 from the National Institues of Health; and Grant-in-Aid Contract M-43 from the State of New Jersey. Recipient of Research Career Award 5-K3-16,749. from the National Institutes of Health.     

Species identity of insect cell lines

Summary Three mosquito cell cultures designated as Suitor's clone ofAedes aegypti, Culiseta inornata, andAedes vexans were shown to be moth by immunological, karyological, and isozyme analyses. The cells reacted with rabbit antimoth serum but not rabbit antimosquito serum. Chromosome analyses indicated Lepidopteran rather than Dipteran morphology, and three isozyme systems were confirmative. Any one of these assays would be sufficient to indicate that contamination had occurred and could be used as a periodic check for identity of cell cultures. Morphology and growth characteristics are also valid criteria to distinguish between these particular orders of insect cells. These studies were supported by Grant CA-04953-12 from the National Cancer Institute; General Research Support Grant FR-5582 from the National Institutes of Health; and Grant-in-Aid Contract M-43 from the State of New Jersey. Recipient of Research Career Award 5-K3-16,749 from the National Institutes of Health.     

Vagotomy and Pyloroplasty for Gastric Ulcer

In a series of 68 patients with gastric ulcer who were treated by vagotomy and pyloroplasty there were no operative deaths, though one patient who received massive transfusions died 14 weeks later from hepatic necrosis probably associated with serum hepatitis. Four patients developed recurrent peptic ulceration requiring further surgical treatment. The clinical condition of all but three of the remainder was satisfactory after an average follow-up period of three years and two months. Of the 14 patients who presented with an acutely bleeding gastric ulcer, one subsequently required gastrectomy for continued bleeding; in all the others the immediate and long-term results were satisfactory. Of 21 patients with a “high” ulcer and 29 with combined gastric and duodenal ulceration one (4·8%) and three (10%) respectively suffered recurrences. We conclude that vagotomy and pyloroplasty is a satisfactory form of treatment for a high or bleeding gastric ulcer, but that for all other gastric ulcers some form of gastric resection is preferable.     

Zonal centrifugation and flotation– fractionation of MSD mutant mouse brain

Zonal centrifuge and flotation–fractionation profile analysis of neonatal mouse brain homogenates in iso-osmotic Ficoll–sucrose density–gradients demonstrates the presence of four light density fractions. In msd neurological mutant mice with a myelin-synthesizing deficiency syndrome, the bands appear to be relatively normal until after the 10th day of postnatal brain development. With the onset of visible neurological symptoms after the 11th day, the four density bands begin to disappear from the zonal profiles and are all but absent at the time of death at about the 21st postnatal day. In normal littermates of the mutants, the bands persist with age and intensify. Although their identities remain unknown, the top three identify by their density with adult myelin and the fourth with the lighter of two adult synaptosome fractions. Mixtures of brain homogenates between mutant and normal littermates give rise to zonal and banding profiles intermediate between the separate profiles but somewhat less than their average in intensity.     

Collagen polysomes: Site of hydroxylation of proline residues

The biosynthesis of collagen on polysomes has been studied by using a newly devised method for obtaining polysomes in high yield from stationary-phase mouse fibroblast (line 3T6; Goldberg &, Green, 1967). These polysomes were completely disaggregated to monosomes by brief exposure to ribonuclease and they lost most of their radioactivity to the top of the sucrose gradients as a result of a 30-minute chase with unlabeled proline. After a ten-minute pulse with [³H]proline, nascent collagen peptides could be identified in these polysomes on sucrose gradients. Most of the proline residues susceptible to hydroxylation by collagen proline hydroxylase were found, in most cases, to be already hydroxylated in these nascent peptides. The nascent nature of these peptides was confirmed by the observation that treatment of the polysomes with RNase transferred the radioactive collagen peptides to the monosome area and these peptides could subsequently be removed to the soluble material at the top of the gradient upon treatment with puromycin. These findings therefore, show clearly that the hydroxylation of proline residues is occurring, in vivo under normal conditions, on nascent collagen chains. In no case was the degree of hydroxylation of the released collagen chains higher than that on the nascent collagen peptides. It seems likely, therefore, that the major site of proline hydroxylation is the nascent collagen peptide.     

Reoccurrence of the microsporidan Perezia pyraustae in the European corn borer, Ostrinia nubilalis, reared on diet containing fumidil B

Fumidil B suppressed the level of Perezia pyraustae infection in the larvae of the European corn borer, Ostrinia nubilalis, while they were feeding on diet containing the drug. After feeding ceased and the gut was emptied of diet containing Fumidil B, the level of infection again increased. Drug levels of 1000 and 1500 ppm in the meridic diet significantly reduced the mean level of P. pyraustae in the adult stage as compared with lower drug levels of 200 and 500 ppm. The level of infection increased throughout the pupal and adult stages, but during the pupal stage it increased much faster in the female than in the male. The rapid increase in the level of infection in the female is discussed in relation to transovariole transmission to the next generation.     

Studies on Nondefective Adenovirus-Simian Virus 40 Hybrid Viruses I. A Newly Characterized Simian Virus 40 Antigen Induced by the Ad2+ND1 Virus

The nondefective adenovirus 2 (Ad2)-simian virus 40 (SV40) hybrid virus, Ad2(+)ND(1), does not induce heat-labile SV40 T antigen but does induce a previously uncharacterized heat-stable SV40 antigen-the SV40 "U" antigen. This antigen is detectable by both immunofluorescence and complement fixation by using sera from hamsters with SV40 tumors. Sera from hamsters bearing SV40 tumors can be divided into two groups, those that react with both SV40 T and U antigens (T(+)U(+) sera) and those that react with SV40 T antigen only (T(+)U(-) sera). SV40 U-specific sera from monkeys immunized with Ad2(+)ND(1)-infected cells do not react with SV40 T antigen by immunofluorescence but do react with an antigen in the nucleus of SV40-transformed cells and with an early, cytosine arabinoside-resistant antigen present in the nucleus of SV40-infected cells. A heat-stable SV40 antigen detectable by complement fixation with T(+)U(+) hamster sera is present in extracts of SV40-induced hamster tumors and in cell packs of SV40-infected or -transformed cells. SV40 U-antigen synthesis by Ad2(+)ND(1) virus is partially sensitive to inhibitors of deoxyribonucleic acid synthesis, whereas U-antigen synthesis by SV40 virus is an early cytosine arabinoside-resistant event. As an early SV40 antigen differing from SV40 T antigen, U antigen may play a role in malignant transformation mediated by SV40.     

An ultraviolet photoreceptor in a Dipteran compound eye

Summary Intracellular recordings of the retinula cell receptor potential to flashes of monochromatic light demonstrate the presence of ultraviolet photoreceptors in the retina of the honeybee-mimic dronefly (Diptera, Syrphidae,Eristalis). Photoreceptors for visible light were also found. Hence, the dronefly has the neural capability for color vision. This is consistent with behavioral studies. Color vision would aid this animal in its natural habit of forraging with the honeybee.Supported by grants AFOSR-71-2112 and NSF GB-30733.