Immunocytochemical localization of a developmentally regulated, isoproterenol-inducible protein (LM protein) in rat submandibular gland

Administration of the beta-adrenergic drug isoproterenol (IPR) produces hyperplastic and hypertrophic enlargements of the submandibular gland of the rat and induces the synthesis of specific proteins in this organ. One of these proteins, the LM (large mobile) protein, was demonstrated immunocytochemically in the submandibular glands of developing untreated and IPR-treated rats. Immunoreactive LM protein was absent in the glands of 20-day-old fetuses and 1- and 2-day-old rats. It was localized in the proacinar and immature acinar cells in the glands of 6- to 21-day-old animals, but it was undetectable at 28 days of age. In the glands of adult rats, secretory granules of the granular convoluted tubule cells showed immunostaining for the LM protein which was also present in trace amounts in the acinar cells. Daily administration of IPR for 5 days to newborn or 8- or 15-day-old rats caused an apparent acceleration of proacinar/acinar cell differentiation, and consequently it increased the frequency of cells immunostained for the LM protein as well as the amount of immunoreactive material in these cells. Thus, the expression of LM protein in the submandibular gland is developmentally regulated, and it is restricted to the stage of differentiation of proacinar cells from terminal tubule cells. IPR is capable of inducing this protein in fully differentiated acinar cells in 3-week-old or older animals.     

Induction of a specific (LM) protein in the submandibular gland of the rat by repeated amputation of the lower incisor teeth

Chronic administration of the beta-adrenergic agonist isoproterenol (IPR) leads to marked hyperplastic/hypertrophic enlargements of the parotid and submandibular glands in rats and mice with concomitant changes in the composition of both the glands and the saliva. Conspicuous among the alterations of the submandibular saliva is the appearance of a 13,000 Mr protein, termed LM (large mobile) protein. Repeated amputation of the lower incisor teeth also causes enlargements of the major salivary glands in rats. In this study, we have compared the enlargements of submandibular glands of rats produced by IPR administration or teeth amputation with respect to the relative levels of the LM protein in gland extracts and saliva. Administration of IPR-HCl (40 mg/kg) twice daily for 5 days or amputation of the lower incisor teeth 3 times a week for 3 weeks resulted in a 2.2-fold increase in the weight of the submandibular gland. Amputation for one week led to a 1.4-fold increase in gland weight. Double immunodiffusion in agar antibodies against the purified LM protein gave a single precipitin line with gland extracts and saliva of IPR-treated and teeth-amputated rats, indicating immunological identity of the reacting antigens. No precipitin lines were seen with gland extracts or saliva of untreated rats. Immunoblots of pooled saliva obtained from IPR-treated or teeth-amputated rats revealed a single protein band of the same electrophoretic mobility in SDS-polyacrylamide gels when stained using anti-LM antibodies. The relative concentrations of LM protein in gland extracts and saliva were measured by a solid-phase enzyme-linked immunoabsorption assay using antibodies against the purified LM protein.(ABSTRACT TRUNCATED AT 250 WORDS)     

Liver function and protein binding in camels

1. Dehydration of camels for 10 days resulted in reduction of liver functions, expressed in longer half life and reduced clearance of bromosulfophthalein (BSP), elevated AST (ALT levels were below the limit of detection of the method) and reduced serum albumin concentrations. 2. Binding of BSP to camel serum proteins by gel permeation chromatography and by equilibrium dialysis showed very strong binding. 3. Binding parameters of various drugs to camels serum by equilibrium dialysis showed close similarities both qualitatively and quantitatively to those of humans. 4. Albumin seems to be the major serum binding protein of BSP.     

A new locus in the phosphate specific transport (PST) region of Escherichia coli

Summary PhoS64 is a mutation in the Phosphate Specific Transport (PST) region on the E. coli chromosome which lacks the periplasmic phosphate binding protein. In contrast to other phoS mutations (which have the same phenotype) it complements the mutations in phoT and pstB. A detailed genetic map of the PST region constructed by three point transductional crosses has revealed that phoS64 is located distally from other phoS mutations. The genetic order obtained was phoS64-phoU35-pstB401-phoT-phoS-ilvC. The data indicate that phoS64 belongs to a different complementation unit in the PST region not known hitherto. We propose to name it phoV.Abbreviations AP alkaline phosphatase - EU enzyme units - Pi inorganic orthophosphate - pNPP paranitrophenyl phosphate - Km kanamycine - Tc^r tetracycline-resistant     

A second site-specific recombination event in the lambdoid bacteriophage HK022

 An in vitro site-specific recombination reaction of the lambdoid phage HK022 has revealed two supercoiled products that proved to be Holliday intermediates. One of them is the Holliday intermediate which has resulted from an attP×attB reaction. The other is an intermediate which has resulted from a recombination reaction between attP and the attL site of the product from the first reaction. The preferential attL×attP over attR×attP reaction was confirmed in vitro and in vivo by challenging attP sites with attL and attR sites. The biased attP×attL over attP×attR reaction in phage HK022 is discussed. Received: 25 April 1996 / Accepted: 11 July 1996     

Guanosine 3',5'-bispyrophosphate (ppGpp) synthesis in cells of Escherichia coli starved for Pi.

Cells of Escherichia coli which enter a phase of starvation for Pi induce the synthesis of the nucleotide guanosine 3',5'-bispyrophosphate (ppGpp). This induction is relA independent but depends on the spoT gene product. A mutant unable to produce ppGpp is impaired in the expression of two genes which belong to the pho regulon, a defect which is dependent on the product of spoT. We suggest that ppGpp is essential for the proper induction of the genes which belong to the pho regulon.     

Site-specific recombination in mammalian cells expressing the Int recombinase of bacteriophage HK022 – Site-specific recombination in mammalian cells promoted by a phage integrase

The int gene of bacteriophage HK022, coding for the integrase protein, was cloned in a mammalian expression vector downstream of the human cytomegalovirus (CMV) promoter. Green monkey kidney cells (COS-1) and mouse embryo fibroblast cells (NIH3T3) transiently transfected with the recombinant plasmid express the integrase protein. Co-transfection of this plasmid with reporter plasmids for site-specific recombination and PCR analyses show that the integrase promotes site-specific integration as well as excision. These reactions occurred without the need to supply integration host factor and excisionase, the accessory proteins that are required for integrase-promoted site-specific recombination in vitro as well as in the natural host Escherichia coli.     

Complementation tests between alkaline phosphatase-constitutive mutants (phoS and phoT) of Escherichia coli.

Complementation tests between phoS and phoT mutations showed that they belong to the same cistron. Homozygosis of a heterozygotic partial diploid resulted from allelic transfer from the chromosome to the F' episome.     

Complementation tests between mutations in the phosphatespecific transport region ofEscherichia coli

Complementation between mutants impaired in inorganic phosphate (Pi) transport via the phosphate-specific transport (PST) system was studied. For that purpose the transport of Pi via the alternative Pi transport (PIT) system was bioenergetically arrested. Complementation was found betweenpstB andphoT mutations, whereas each of these mutations failed to complement with aphoS mutation. The data obtained confirm previous studies in which the inducibility of alkaline phosphatase was used to determine complementation and indicated a polar effect of thephoS mutation onpstB andphoT.