Formation of a thiamine artifact during chromatography: A single column procedure for the separation of thiamine and the thiamine mono-, di-, and triphosphate esters

Chromatography of a preparation of [¹⁴C]thiamine pyrophosphate (thiamine-PP) on Dowex 1X8 in the formate form produced an unexpected peak, X-1, which was eluted just prior to the thiamine-PP peak. An ammonium formate, pH 4.5, gradient was the eluant. Rechromatography of either peak X-1 or thiamine-PP produced the same two peaks. The radioactive specific activity per micromole labile phosphate was the same for the two peaks. Peak X-1 appears to be a thiamine compound formed in the presence of formate solutions. This procedural artifact was circumvented by the substitution of acetate for formate. By varying the pH as well as the ionic strength a single column procedure has been developed that separates thiamine and the three phosphate esters quantitatively in micromolar amounts.     

Comparison of antibody production against aflatoxin B1 in goats and rabbits.

Bacteria were found that are capable of producing good yields of β-amylase in unrefined media. The culture filtrates are free of α-amylase and isoamylase.     

SOLUBLE AND MEMBRANE LECTIN-BINDING GLYCOPROTEINS OF THE CHROMAFFIN GRANULE

Abstract— Fluorescein isothiocyanate-labelled lectins were used to identify lectin-binding glycoproteins of the chromaffin granule after electrophoresis of the membrane and soluble granule proteins on sodium dodecyl sulphate polyacrylamide slab gels. The glycoprotein nature of all lectin-binding bands was confirmed by staining the gels for carbohydrates, and the specificity of the lectin-binding was demonstrated by hapten sugar inhibition of binding. In samples of granule membrane proteins reduced with dithiothreitol 10 concanavalin A (Con A), 5 wheat germ agglutinin, 8 Ricinus communis agglutinin-60, and 7 Ricinus communis agglutinin-120 (RCA-120) binding glycoproteins were identified. Molecular weights of these glycoproteins varied from 20,000 to 200,000 daltons. All but two of the Con A-binding bands and one of the RCA-120 binding bands appeared to react with more than one lectin, suggesting possible carbohydrate heterogeneity in these membrane glycoproteins. The band identified as dopamine β-hydroxylase reacted most intensely with all four lectin tested, and in the soluble core material this enzyme was the sole significant lectin binding glycoprotein.     

Seasonal species composition of barnacle larvae (Cirripedia: Thoracica) in Rhode Island waters, 1977-1978

Cirripede larvae can occur year-round in temperate and tropicalwaters, often in significant numbers, yet the species compositionof the ‘Balanus sp.’ component is rardy studied.Weekly plankton samples were analyzed qualitatively for larvalcirripede species and stage over a year (1977–1978) attwo Rhode Island stations. Six species of larvae were foundin Lower Narragansett Bay (30° salinity). Semibalanus balanoidesand Balanus balanus have a single winter brood. S. balanoidesis the predominant winter breeder with a minor release of naupliiin early December and major release in March followed by cypridsin mid-April.B. balanus populations release all larvae in Marchwith cyprids in mid-April. Balanus crenatus is mainly a winterbreeder, but has multiple broods; it does not breed in July-Septemberwhen the water temperature is above 18°C. Balanus venustusis the predominant summer breeder, and larvae were observedfrom May through December (water >8°C). Larvae of Chthamalusfragilis and Balanus eburneus occur in low numbers from May-October.At the Pettaquamscutt River site (12 salinity), Balanus improvisuslarvae predominate and early stage nauplii (I-II) occur in samplesyear round (0–27°C). Two peaks of later stage naupliiand cyprids occur in late spring (May) and early winter (Nov.-Jan.).Continued temperatures bdow 5°C or above 20°C appearto inhibit larval development. Comparison of results with existing literature reveals severalsignificant findings. The bimodal rdease of S. balanoides larvaeis unusual and may be in response to the phytoplankton dynamicsof the year; however, the existence of distinct races of S.balanoides may also be a factor. Larvae of B. venustus predominatein the lower bay during the summer, yet this species is unreportedin past studies. B. improvisus nauplii are more cold tolerantthan previously reported. Comparison of findings with reportedbreeding patterns in Florida indicate significant differencesin temperature responses between northern and southern populations. ¹Contribution No. 188 from EPA Environmental Research Laboratory,South Ferry Rd., Narragansett, RI 02882     

Growth of dissociated rat cerebellar cells using serum-free supplemented media and varied transferrin concentrations

Dissociated neonatal rat cerebellar cells were grown on medium supplemented with 10% horse serum (HS) and compared with those grown using a serum-free supplemented (SFS) medium, modified from Bottenstein and Sato (1979). containing insulin, transferrin, progesterone, putrescine, and selenium (after an initial 24 hr in 10% horse serum). Cells survived for several weeks using either medium. Cells grown in SFS had higher levels of GABA uptake than cells grown in HS. Cellular morphology and the proportion of neurons to glial cells were similar under the two conditions. Transferrin concentrations of 0.5, 10, and 100 µg/ml were tested. Neither neuronal nor glial cells were sensitive to this 200-fold variation. The SFS medium supports survival and maturation of both neurons and glial cells from rat cerebellum. However, the medium is not completely defined since (1) one day of serum is still required and (2) the heterogeneous cell population is undoubtedly conditioning the medium to some extent.This work was supported in part by grants from the Scottish Rite Schizophrenia Research Program, N.M.J., USA, and by Biomedical Research Grant S 07-RR5394, from the National Institutes of Health, PHS/DHHS.     

Selenocystine-resistant mutants of Histoplasma capsulatum

Cysteine metabolism has been thought to be important to the phenomenon of dimorphism inHistoplasma capsulatum. We sought mutants with genetic blocks in the metabolism of cysteine by selection of colonies resistant to the toxic analogue, selenocystine. The 22 resistant strains thus obtained were all deficient in uptake of cystine from the surrounding medium but were normally able to convert from mycelium to yeast and back again. Furthermore, they had normal quantities of NADH-dependent cystine reductase when this enzyme was measured. We conclude that mutants defective in cystine uptake can be readily obtained by selection of colonies resistant to selenocystine, and that a lesion in cystine-uptake does not appear to affect the phenomenon of dimorphism in this organism.Preliminary reports of this work were presented at the Second International Congress of Mycology, Tampa, 1977 and at the first International Conference on Histoplasmosis, Atlanta, 1978.     

Two cases of trisomy 21 and one XXY case with atypical clinical features

Summary Three patients with mental retardation and multiple congenital abnormalities are described.Although their clinical appearance was not suggestive of Down's syndrome, chromosome studies showed a non-disjunctional trisomy 21 in two of the patients. The third case had an unsuspected XXY karyotype.     

Role of adrenergic innervation in experimental renal hypertension

Renal hypertension was induced by ligation of the aorta between renal arteries in rats sympathectomized with 6-hydroxydopamine. In the early phase, equally severe hypertension developed in the denervated group as compared to innervated controls. Later, blood pressure was lower in the denervated rats. Initially, increases in plasma renin were seen in both groups; the levels, however, were markedly lower in the denervated rats. Later, the renin levels were similar and not different from baseline. It is concluded that adrenergic neural activity is not essential in the development of renal hypertension; the maintenance of the chronic state, however, depends in part on adrenergic innervation.