Ethoxyformylation of histidine residues in equine growth hormone

Reactivity of histidine residues in equine growth hormone to ethoxyformic anhydride was studied. The existence of two kinetically different sets was demonstrated: one of them including only the slow reacting histidine 169 (k = 0.164 min-1) and the other containing fast reacting histidines 19 and 21 (k = 0.892 min-1). A correlation between the decrease in the capacity to compete with 125I-labeled hormone for rat liver binding sites and the degree of ethoxyformylation of the fast group was found. Circular dichroism studies indicated no significant conformational changes in the protein with all three residues modified. These results fully agree with those obtained for bovine growth hormone which is further evidence supporting the vinculation of histidines 19 and/or 21 with the binding site of these hormones to their specific receptors.     

The effect of water activity and pH on the production of mycotoxins by fungi growing on a bread analogue

Mycotoxin production by various toxigenic fungi, growing on a bread analogue, was investigated at various water activities (a_w) and pH combinations. Citrinin, ochratoxin A and sterigmatocystin could be detected at a_w > 0·80, while patulin was only observed at a_w= 0·95. These results show that some toxins may be produced at lower water activities than have been reported on synthetic media and suggest that, where possible, natural substrates should be used to investigate factors affect-ing mycotoxin production in foodstuffs.     

Keratinocyte growth-promoting activity from human placenta

Extracts of term human placenta were tested for enhancement of proliferative growth of primary cultures of human keratinocytes. Saline extracts or supernatants from homogenates were dialyzed extensively, lyophilized, and tested in subcultures of keratinocytes in MCDB 153 medium with 0.1 mM Ca++ containing only defined supplements (insulin, hydrocortisone, transferrin, ethanolamine, phosphoethanolamine). Cells plated in the absence of EGF at moderately high densities (1000-3000 cells per cm2) formed colonies and grew in the presence of placental extract at 25-500 micrograms/ml. Extracts of cord serum or maternal serum were inactive, suggesting that the activity is derived from placental tissue. The activity is not EGF, since the activity in the placental extract, unlike EGF, did not promote growth at low cell density, was synergistic with EGF under some conditions, and did not produce changes in colonial morphology which occurred in the presence of EGF. Unlike keratinocyte growth-promoting activity in bovine hypothalamic extract, the activity is non-dialyzable and is destroyed at 100 degrees C. Placental extract could not replace any of the defined components of the medium and is therefore distinct from them. The presence of activity in the placenta with distinctive properties suggests that this is a previously undescribed material with growth-promoting properties for epithelium.     

Biosynthesis of ferredoxin in Chlamydomonas reinhardii

The selective action of the antibiotics chloramphenicol and cycloheximide on the synthesis of ferredoxin in liquid cultures of Chlamydomonas reinhardii was studied. Highly specific antibodies raised against Chlamydomonas ferredoxin were used to determine the in vivo synthesis of apoferredoxin and conversion into native protein. The results indicate that 80S ribosomes are involved in the synthesis. Chlamydomonas cells growing in the absence of iron did not synthesize immunologically detectable amounts of ferredoxin. We suggest that this is based upon feed-back inhibition of apoferredoxin synthesis at the translational level.Abbreviations CAP chloramphenicol - CHI cycloheximide - IgG Immunoglobulin G - PBS 140.4 mM NaCl. 9 mM Na₂HPO₄, 1.3 mM NaH₂PO₄ (pH 74) - SDS sodium dodecvl sulphate - Fd Ferredoxin - apoFd Apoferredoxin - CM-Fd Scarboxymethylated Fd - TCA-Fd Fd treated with trichloroacetic acid     

Polyacrylamide slab gel electrophoresis: an improved design

An apparatus for polyacrylamide slab gel electrophoresis is decribed which combines all parts into one integral unit. It eliminates several steps in the process of sealing, pouring, and setting the gels. Construction is easy with modest workshop facilities and the design easily adapted to suit most requirements. The apparatus provides a high degree of versatility and is suitable for use with many slab gel electrophoretic techniques.     

An overview of some mechanisms of bacterial pathogenesis

The adherence of microorganisms to host surfaces is highly specific, and in many cases, essential for subsequent pathogenetic events to occur. A dynamic process leading to increased mucosal adherence of gram-negative bacilli to epithelial cell receptors in the oral cavity appears to be the initial step in the development of pneumonia. In infectious processes secondary to Streptococcus pneumoniae, adherence may also play a role in specific syndromes. In many cases, however, colonization of oropharyngeal mucus itself, the presence of capsular polysaccharide, and the release of various cell wall components appear to interact to cause clinical disease. In Neisseria gonorrhoeae infections, adherence is all important and is mediated by a number of cell surface structures. These have been studied extensively. Many of these structures, such as pili and protein II, exhibit great variability both between strains and in the same organism at different stages of infection. Others, such as protein I, are more constant. This information has been used in the production of specific vaccines to more preserved structures to inhibit adherence. These will be tested in the near future. It is our view that a better understanding of the many forms of bacterial adherence will be the key to our designing more effective strategies to detect early infection and to intervene more decisively to limit its spread.