Cloning and Characterization of Two Human G Protein-Coupled Receptor Genes (GPR38 and GPR39) Related to the Growth Hormone Secretagogue and Neurotensin Receptors

The recent cloning of a growth hormone secretagogue receptor (GHS-R) from human pituitary gland and brain identified a third G protein-coupled receptor (GPC-R) involved in the control of growth hormone release. The nucleotide sequence of the GHS-R is most closely related to the neurotensin receptor-1 (NT-R1) (35% overall protein identity). Two human GPC-Rs related to both the type 1a GHS-R and NT-Rs were cloned and characterized. Hybridization at low posthybridizational stringency with restriction enzyme-digested human genomic DNA resulted in the identification of a genomic clone encoding a first GHS-R/NT-R family member (GPR38). A cDNA clone was identified encoding a second GHS-R-related gene (GPR39). GPR38 and GPR39 share significant amino acid sequence identity with the GHS-R and NT-Rs 1 and 2. An acidic residue (E124) in TM-3, essential for the binding and activation of the GHS-R by structurally dissimilar GHSs, was conserved in GPR38 and GPR39. GPR38 is encoded by a single gene expressed in thyroid gland, stomach, and bone marrow. GPR39 is encoded by a highly conserved single-copy gene, expressed in brain and other peripheral tissues. Fluorescencein situhybridization localized the genes for GPR38 and GPR39 to separate chromosomes, distinct from the gene encoding the GHS-R and NT-R type 1. The ligand-binding and functional properties of GPR38 and GPR39 remain to be determined.     

The metabolism of glucose by heterocysts and vegetative cells of Anabaena cylindrica

Whole filaments of autotrophically grown Anabaena cylindrica and heterocysts isolated from them will assimilate and metabolise exogenous glucose. Radiorespirometric experiments suggest the operation of the pentose phosphate pathway. Glucose-6-phosphate and 6-phosphogluconate dehydrogenase are present in heterocysts at 6–8 times the levels found in vegetative cells whereas enzymes of the reductive pentose phosphate and glycolytic pathways are barely or not detectable. Glucose-6-phosphate dehydrogenase in vegetative cells, but not in heterocysts is subject to inhibition by ribulose diphosphate.     

Efficient soybean transformation using hygromycin B selection in the cotyledonary-node method

The efficiency of soybean [Glycine max (L.) Merrill] transformation was significantly increased from an average of 0.7% to 16.4% by combining strategies to enhance Agrobacterium tumefaciens-mediated T-DNA delivery into cotyledonary-node cells with the development of a rapid, efficient selection protocol based on hygromycin B. Wounded cotyledonary-node explants were inoculated with A. tumefaciens carrying either a standard-binary or super-binary plasmid and co-cultivated in the presence of mixtures of the thiol compounds, L-cysteine, dithiothreitol, and sodium thiosulfate. Transformed shoots began elongating only 8 weeks after co-cultivation. Southern analysis confirmed integration of the T-DNA into genomic DNA and revealed no correlation between the complexity of the integration pattern and thiol treatment applied at co-cultivation. All T(0) plants were fertile and the majority of the lines transmitted the beta-glucuronidase (GUS) phenotype in 3:1 or 15:1 ratios to their progenies.     

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A synthetic analogue of Escherichia coli lipoprotein, tripalmitoyl pentapeptide, constitutes a potent immune adjuvant

Lipoprotein from the outer membrane of Escherichia coli and other Enterobacteriaceae constitutes a potent B lymphocyte mitogen and polyclonal activator in various species. Tripalmitoyl pentapeptide (S-(2,3-bis-(palmitoyloxy)-(2RS)-propyl)-N-palmitoyl-(R)-cysteinyl -(S)-seryl-(S)-seryl-(S)-asparaginyl-(S)-alanine) is a synthetic analogue of the N-terminal part of lipoprotein and has, in all assays tested, a biologic activity similar to native lipoprotein. It also exhibits a strong adjuvant activity in vitro: In the presence of 3.3 to 33.3 micrograms/ml of tripalmitoyl pentapeptide, the stimulation of the primary antibody response toward underivatized sheep red blood cells (SRBC) and toward trinitrophenylated (TNP-) SRBC was markedly enhanced, as measured by a direct hemolytic plaque assay. At optimal mitogen- and antigen-doses, plaque formation was increased up to 100-fold, and at suboptimal doses (0.03 to 0.3 microgram/ml) a 10- to 60-fold increase of plaque numbers was achieved. In the presence of tripalmitoyl pentapeptide, the antigen-specific IgM response was increased about sevenfold and the IgG response was augmented about 10-fold, as measured by ELISA. Similarly, in the secondary in vitro response to TNP-SRBC, a 7 to 10-fold enhancement of the antibody titer was obtained in the presence of the adjuvant. The application of tripalmitoyl pentapeptide and antigen had to occur concurrently in order to achieve a strong adjuvant effect. Addition of tripalmitoyl pentapeptide to the cell cultures 1 day after or 1 day before antigen application had no significant positive effect, and in several instances a decrease in antibody production was found. Thus, tripalmitoyl pentapeptide, a well-characterized synthetic product available in major amounts, constitutes a potent immune adjuvant for potential animal and clinical use.     

Neuromuscular electrical stimulation increases muscle protein synthesis in elderly type 2 diabetic men

Physical activity is required to attenuate the loss of skeletal muscle mass with aging. Short periods of muscle disuse, due to sickness or hospitalization, reduce muscle protein synthesis rates, resulting in rapid muscle loss. The present study investigates the capacity of neuromuscular electrical stimulation (NMES) to increase in vivo skeletal muscle protein synthesis rates in older type 2 diabetes patients. Six elderly type 2 diabetic men (70 ± 2 yr) were subjected to 60 min of one-legged NMES. Continuous infusions with l-[ring-(13)C(6)]phenylalanine were applied, with blood and muscle samples being collected regularly to assess muscle protein synthesis rates in both the stimulated (STIM) and nonstimulated control (CON) leg during 4 h of recovery after NMES. Furthermore, mRNA expression of key genes implicated in the regulation of muscle mass were measured over time in the STIM and CON leg. Muscle protein synthesis rates were greater in the STIM compared with the CON leg during recovery from NMES (0.057 ± 0.008 vs. 0.045 ± 0.008%/h, respectively, P < 0.01). Skeletal muscle myostatin mRNA expression in the STIM leg tended to increase immediately following NMES compared with the CON leg (1.63- vs. 1.00-fold, respectively, P = 0.07) but strongly declined after 2 and 4 h of recovery in the STIM leg only. In conclusion, this is the first study to show that NMES directly stimulates skeletal muscle protein synthesis rates in vivo in humans. NMES likely represents an effective interventional strategy to attenuate muscle loss in elderly individuals during bed rest and/or in other disuse states.     

Ethnic patterns of cytologic abnormalities in cervical smears in suriname, a high-risk area for cervical cancer

OBJECTIVE: To determine the prevalence of cytologic abnormalities in cervical smears from women attending the first organized screening program in Suriname and to compare the prevalences in 4 Surinamese ethnicities with different cervical carcinoma incidences. STUDY DESIGN: Cervical scrapes were taken from women with 4 different ethnicities: Maroons, Amerindians, Javanese and Hindustani. Papanicolaou staining and cytologic screening were performed on 807 cervical smears. RESULTS Cervical cytologic abnormalities were seen in 13.4%, of which 8.1% (62 of 764) had atypical changes, 2.6% (20 of 764) had mild and 2.6% (20 of 764) had moderate and severe dysplasia/carcinoma in situ (CIS). The cytologic abnormalities varied between the ethnicities: 42.1% (83 of 197) in the Maroons and 2.3% (4 of 176), 5.0% (9 of 183) and 3.0% (6 of 208) in the Javanese, Amerindians, and Hindustani, respectively. CONCLUSION: The high prevalence of moderate and severe dysplasia/CIS in all ethnicities correlates with the high cervical carcinoma incidence in Suriname. A significantly higher prevalence of mild abnormalities in the Maroons was observed; it did not reflect the relatively low cervical cancer incidence in this ethnicity. However, this can be explained by the possibility that these women have a different sexual lifestyle, leading to a higher prevalence of     

Synthesis and cannabinoid-1 receptor binding affinity of conformationally constrained analogs of taranabant

The design, synthesis, and binding activity of ring constrained analogs of the acyclic cannabinoid-1 receptor (CB1R) inverse agonist taranabant 1 are described. The initial inspiration for these taranabant derivatives was its conformation 1a, determined by ¹H NMR, X-ray, and molecular modeling. The constrained analogs were all much less potent than their acyclic parent structure. The results obtained are discussed in the context of a predicted binding of 1 to a homology model of CB1R.