Age- and diapause-related acid and alkaline phosphatase activities in the intestine and malpighian tubules of the Colorado potato beetle, Leptinotarsa decemlineata (Say).

Specific activities for soluble (s) and membrane (m)-bound acid (ACP) and alkaline phosphatases (ALP) were determined in the midgut, hindgut, and Malpighian tubules for developing, prediapausing, and diapausing adult Colorado potato beetles, Leptinotarsa decemlineata (Say). High ACP activities were found in the hindgut and Malpighian tubules while high ALP activities were found in the Malpighian tubules. Variation in both ACP and ALP activities in each tissue reflects fluctuation in protein synthesis and secretion involved with digestion, excretion, and other unknown functions. Phosphatase activities in the tissues examined show the dynamic nature of diapause in this insect. Diapausing beetles showed increases in phosphatase activity after hormone treatments. JHA treatments increased s-ACP and m-ACP activities in all tissues but 20-HE did not increase activity in any tissue. Allatotropin tended to mimic the effects of JHA treatment. The s-ALP activity was also increased in all tissues whereas m-ALP was increased in the midgut and hindgut by JHA treatment. Malpighian tubule m-ALP activity was only increased by 20-HE treatments. Allatotropin was not as effective in increasing ALP activities as it was with ACP activities.

Effects of tissue extracts on oviduct contraction in the migratory grasshopper, Melanoplus sanguinipes.

The isolated oviduct of the migratory grasshopper, Melanoplus sanguinipes, shows a myogenic, rhythmic pattern of contraction. The pattern of contraction can be modified by treatment with hemolymph or extracts from a variety of tissues from animals of differing age, sex, and mated status. Extracts of oviducts and ventral nerve cord (VNC), as well as hemolymph, from both virgin and mated females were almost always stimulating, though their effect on frequency, amplitude and/or tonus varied. In contrast, whereas extracts of spermatheca and brain from mated females were stimulating, extracts of these tissues from virgin females inhibited contraction. All male material tested (long hyaline tubules (LHT), the male accessory gland complex less the LHT, testes, brain, VNC and hemolymph) stimulated oviduct contraction in a dose-dependent manner, usually enhancing at least two of frequency, amplitude and tonus. However, oviposition-stimulating protein, a purified product of the LHT, provoked only an increase in frequency when applied to the oviduct. LHT extract modulated the effects of virgin female tissue extracts, always in a stimulatory manner.

Purification and characterization of an oviposition-stimulating protein of the long hyaline tubules in the male migratory grasshopper, Melanoplus sanguinipes.

An oviposition-stimulating protein (OSP) was isolated and purified from the long hyaline tubules of the male accessory gland complex in the migratory grasshopper, Melanoplus sanguinipes. Gel filtration of the native OSP, using Sephadex G-100, indicates its molecular weight to be about 60000Da with the oviposition-stimulating activity while sodium dodecyl sulphate-polyacrylamide gel electrophoresis shows that the OSP comprises two subunits, each with a molecular weight of 30000Da. The purified OSP appears as a single symmetric peak on fast performance liquid chromatography using Mono Q. Isoelectric focusing of the OSP indicates an apparent pI of 5.5. Injection of the OSP induces oviposition in about 70% of ovulated virgin females within 48h. Stimulation of oviposition can be blocked by a polyclonal antibody raised against the OSP 30000Da subunits. Amino acid analysis of the dimer and its subunits shows a comparatively high content of aspartic acid/asparagine (14.8%) as well as leucine (12.2%) and glutamic acid/glutamine (12.0%). The N-terminal 21 amino acid sequence of the OSP shows little similarity to known peptides. Immunoreactivity with the anti-OSP antibody was observed in the viscous secretion, spermatheca, and the egg-pod froth of mated females, confirming transfer of the OSP from male to female during copulation.

Isolation, identification, and synthesis of Mas-MG-MT I, a novel peptide from the larval midgut of Manduca sexta (Lepidoptera: Sphingidae).

A five-residue myotropic peptide, Manduca sexta midgut myotropin I (Mas-MG-MT I), was isolated from an extract of 800 midguts of fifth instar larvae of the tobacco hornworm, Manduca sexta. It was purified by reverse phase and normal phase HPLC. Myotropic activity was screened by a heterologous Locusta migratoria oviduct bioassay. Sequence analysis, amino acid composition analysis, and comparison of candidate synthetic peptides in the amide and acid form revealed the following primary structure: Ala-Glu-Pro-Tyr-Thr-NH2. This is the first fully identified peptide isolated directly from the midgut of an insect species. Few significant sequence homologies with known vertebrate and invertebrate peptides have been found.

New type of X-linked progressive muscular dystrophy involving shoulder girdle and back.

A new type of X-linked muscular dystrophy is described in a family in which 7 men had boyhood onset of progressive dystrophy involving muscles of the shoulder and back but not the calves or face. The scapula-back muscles are affected, but the calf muscles are normal. All patients are still able to walk. The oldest patient is now 37 years old. The muscular dystrophy has been specified by electromyography, pathologic tissue microscopic examination, electron microscopic study, and elevated CK. This type of muscular dystrophy has not been reported previously.