Barbara E. C. Banks

New methods of isolating bee venom peptides

Abstract The three major peptides of bee venom, melittin, apamin, and peptide 401 (MCD peptide), can be readily separated on columns of Heparin Sepharose CL-6B at pH 6.8 (phosphate buffer) using a linear salt gradient. Contamination by melittin of the other two components can be assessed by fluorescence spectrophotometry.

Immunological properties of nerve growth factors

Antisera were raised against nerve growth factors isolated from mouse salivary gland and five venoms representative of the three main families of poisonous snakes. Immunochemical cross-reactivity was assessed from the ability of the antisera to inhibit the biological activities of the heterologous antigens. The high and low molecular weight forms of the salivary gland factors were found to be immunologically identical but distinct from the snake venom factors. The snake venom factors, on the other hand, were immunologically closely related to each other but not identical.

Nerve growth factor has anti-inflammatory activity in the rat hindpaw oedema test

Nerve growth factor (NGF) affects the development of sympathetic neurons in neonatal rodents and, at very much lower levels, stimulates fibre growth from certain neurones in culture. At levels comparable to those used in culture and found in mammalian sera, NGF has recently been shown to suppress the hindpaw oedema induced in adult rats by injection of carrageenin. On a weight basis, NGF is ten times more active in this test than dexamethasone, and a thousand times more active than the non-steroidal anti-inflammatory drug, indomethacin. It is equally effective given locally and systemically.

The preparation of nerve growth factor from the prostate of the guinea-pig and isolation of immunogenically pure material from the mouse submandibular gland

It was recently discovered [ 1] in these laboratories that the prostate of the guinea-pig is (like the submandibular gland of the adult male mouse) a rich source of nerve growth factor (NGF). We now report, by a modification of the method in [2], the total purification of the factor from this source and describe some of its properties. A similar purification method can also be used to prepare the p-subunit of NGF from mouse submandibular gland free of immunogenic contaminants. We have shown [3] that such contaminants are extremely widespread and may seriously interfere with the interpretation of biochemical and biological experiments.

The effects of postganglionic axotomy and nerve growth factor on the superior cervical ganglia of developing mice

SummarySectioning of the two major outflows from the superior cervical ganglia in two week mice results in a marked drop in the number of neurons within one week of operation and a smaller drop over the following two weeks. In animals receiving daily injections of nerve growth factor (NGF), the effect of axotomy is modified. One week after axotomy, the number of neurons in the axotomized ganglia is approxima the same in NGF treated animals as in the control, sham operated ganglia. Over the next two weeks, however, the cell death that results from axotomy is no longer prevented by treatment with NGF. The normal, hyperplastic response to NGF appears to occur independently of the cell reaction caused by axotomy.

A comparative study of nerve growth factors from snake venoms

Abstract 1. 1. Methods for the partial purification of nerve growth factor from five different snake venoms are described. 2. 2. The physico-chemical properties of the different proteins are compared and the difficulties involved in the isolation of the factor from snake venoms are discussed.

Anti-inflammatory activity of bee venom peptide 401 (mast cell degranulating peptide) and compound 48/80 results from mast cell degranulation in vivo.

1 The relationship between the anti‐inflammatory activity of the bee venom peptide 401 in the carrageenin‐induced oedema of the rat hind paw and its mast cell degranulating activity has been reinvestigated. 2 Mast cell degranulation caused by compound 48/80 (10 mg kg−1) or by allergen challenge in rats sensitized to Nippostrongylus brasiliensis also suppressed rat hind paw oedema in the same test. 3 The anti‐inflammatory activities of peptide 401 and compound 48/80 were partially suppressed by pretreatment of rats with mepyramine and methysergide, at doses (2.5 mg kg−1) that completely suppressed skin reactions to these mast cell‐derived amines. Pretreatment of rats with compound 48/80 also suppressed the apparent anti‐inflammatory actions of peptide 401 and of compound 48/80. 4 Injection of peptide 401 together with carrageenin increased the inflammatory response in the rat hind paw. 5 The anti‐inflammatory activity of peptide 401 and of compound 48/80 in the carrageenin‐induced swelling of the rat hind paw arises from mast cell degranulation in vivo.

A program to compute the sizes and numbers of spherical bodies from observations made on tissue sections

SummaryA computer program, written in standard Fortran, is given to facilitate the computation of the sizes and correct distribution of spheres from the number and size of the circular profiles observed in tissue sections. The method is particularly valuable in cases where it is necessary to determine absolute numbers of cells or subcellular fragments where the objects of interest are not of uniform size. The method gives true cell numbers and the distribution of the spherical diameters.

The isolation and identification of noradrenaline and dopamine from the venom of the honey bee, Apis mellifica

Abstract The low molecular weight components of the venom of the common European honey bee were separated by dialysis and fractionated by gel filtration and ion exchange chromatography. Two fractions showing anti-inflammatory activity in the carrageenin-induced rat paw oedema test were detected. One was identified as the Mast Cell Degranulating Peptide, otherwise known as Peptide 401. The other gave, after conversion to the pentafluoropropionate derivative, two volatile compounds which were shown by combined gas-liquid chromatography and mass spectrometry, to be noradrenaline and dopamine.

Differing effects on superior cervical ganglia in neonatal mice produced by antisera to nerve growth factors from mice and snakes

Abstract Rabbit or horse antisera to Nerve Growth Factors from mouse salivary glands and the venoms of five snakes were injected into neonatal mice. The mice were killed 10 days later and the superior cervical ganglia removed, weighed and examined histologically. Treatment with antisera to the snake Nerve Growth Factors had no effect on ganglion weight, maximum neuronal density or mean neurone diameter. This was true even for the one antiserum that in vitro showed a weak cross-reactivity with the mouse antigen. In contrast, treatment with the antiserum to mouse Nerve Growth Factor produced a partial destruction of the superior cervical ganglia (immunosympathectomy). The weights of the ganglia in animals treated for the first 5 days post partum with increasing volumes (a total of 0.5 ml) of antiserum to the mouse factor fell by some 80% the total neurone number by approx 50% the maximum neurone density by 40% and the mean neurone dia. by 17%. The effect was found to be dose dependent. It is considered that caution is required in extrapolating results from in vitro studies on Nerve Growth Factor to the situation obtaining in vivo and that the inability of snake antisera to produce immunosympathectomy in neonatal mice may result from differences in the antigenic determinants of the mouse and snake Nerve Growth Factors. The marked effect of antiserum to the mouse salivary gland factor in neonatal mice reported by earlier workers has been confirmed. No single explanation, however, can be given for the differences in the reduction in neurone numbers found in the present and previous studies. Furthermore, it is concluded that neither the previous nor the present studies afford evidence that the antiserum directly causes neuronal death.