Consequences of Cis-amide Bond Simulation in Opioid Peptides

Summary Six analogs of leucine-enkephalin were synthesized in which a 1,5-disubstituted tetrazole ring was incorporated in order to lock selected peptide bonds in cis geometry. The obtained compounds were examined based on their biological effects in vivo and in vitro. Only one analog was completely inactive in binding assays being very weakly active in the antinociceptive test. The remaining five compounds displayed at least weak receptor affinity and in vivo activity.     

Effect of glycosylation inhibitors on binding properties of the Fc gamma receptors from guinea pig peritoneal macrophages.

Guinea pig peritoneal macrophages possess on their surfaces receptors for the Fc region of IgG immunoglobulins (Fc gamma R). The cells treated with the glycosylation inhibitors tunicamycin or monensin interacted with IgG with a higher affinity and showed a lower number of IgG-binding sites in comparison with control cells. This indicates that the interaction of IgG with the macrophage Fc gamma receptor depends on the degree of glycosylation of the cell surface glycoconjugates and that glycosylation of the macrophage Fc gamma receptor is important for the expression of the mature form of the receptor.     

Spermatogenic cells of the prepuberal mouse: isolation and morphological characterization

A procedure is described which permits the isolation from the prepuberal mouse testis of highly purified populations of primitive type A spermatogonia, type A spermatogonia, type B spermatogonia, preleptotene primary spermatocytes, leptotene and zygotene primary spermatocytes, pachytene primary spermatocytes and Sertoli cells. The successful isolation of these prepuberal cell types was accomplished by: (a) defining distinctive morphological characteristics of the cells, (b) determining the temporal appearance of spermatogenic cells during prepuberal development, (c) isolating purified seminiferous cords, after dissociation of the testis with collagenase, (d) separating the trypsin-dispersed seminiferous cells by sedimentation velocity at unit gravity, and (e) assessing the identity and purity of the isolated cell types by microscopy. The seminiferous epithelium from day 6 animals contains only primitive type A spermatogonia and Sertoli cells. Type A and type B spermatogonia are present by day 8. At day 10, meiotic prophase is initiated, with the germ cells reaching the early and late pachytene stages by 14 and 18, respectively. Secondary spermatocytes and haploid spermatids appear throughout this developmental period. The purity and optimum day for the recovery of specific cell types are as follows: day 6, Sertoli cells (purity>99 percent) and primitive type A spermatogonia (90 percent); day 8, type A spermatogonia (91 percent) and type B spermatogonia (76 percent); day 18, preleptotene spermatocytes (93 percent), leptotene/zygotene spermatocytes (52 percent), and pachytene spermatocytes (89 percent), leptotene/zygotene spermatocytes (52 percent), and pachytene spermatocytes (89 percent).     

A general method for preparation of N‐Boc‐protected or N‐Fmoc‐protected α,β‐didehydropeptide building blocks and their use in the solid‐phase peptide synthesis

N‐(tert‐butyloxycarbonyl) or N‐(9‐fluorenylmethoxycarbonyl) dipeptides with C‐terminal (Z)‐α,β‐didehydrophenylalanine (?^ZPhe), (Z)‐α,β‐didehydrotyrosine (?^ZTyr), (Z)‐α,β‐didehydrotryptophan (?^ZTrp), (Z)‐α,β‐didehydromethionine (?^ZMet), (Z)‐α,β‐didehydroleucine (?^ZLeu), and (Z/E)‐α,β‐didehydroisoleucine (?^Z/EIle) were synthesised from their saturated analogues via oxidation of intermediate 2,5‐disubstituted‐oxazol‐5‐(4H)‐ones (also known as azlactones) with pyridinium tribromide followed by opening of the produced unsaturated oxazol‐5‐(4H)‐one derivatives in organic‐aqueous solution with a catalytic amount of trifluoroacetic acid or by a basic hydrolysis. In all cases, a very strong preference for Z isomers of α,β‐didehydro‐α‐amino acid residues was observed except of the ΔIle, which was obtained as the equimolar mixture of Z and E isomers. Reasons for the (Z)‐stereoselectivity and the increased stability of the aromatic α,β‐didehydro‐α‐amino acid residue oxazol‐5‐(4H)‐ones over the corresponding aliphatic ones are also discussed. It is the first use of such a procedure to synthesise peptides with the C‐terminal unsaturated residues and a peptide with 2 consecutive ΔPhe residues. This approach is very effective especially in the synthesis of peptides with aliphatic α,β‐didehydro‐α‐amino acid residues that are difficult to obtain by other methods. It allowed the first synthesis of the ?Met residue. It is also more cost‐effective and less laborious than other synthesis protocols. The dipeptide building blocks obtained were used in the solid‐phase synthesis of model peptides on a polystyrene‐based solid support. Peptides containing aromatic α,β‐didehydro‐α‐amino acid residues were obtained with PyBOP or TBTU as a coupling agent with good yields and purities. In the case of aliphatic α,β‐didehydro‐α‐amino acid residues, a good efficiency was achieved only with DPPA as a coupling agent.     

Mitochondria and the dynamic control of stem cell homeostasis

The maintenance of cellular identity requires continuous adaptation to environmental changes. This process is particularly critical for stem cells, which need to preserve their differentiation potential over time. Among the mechanisms responsible for regulating cellular homeostatic responses, mitochondria are emerging as key players. Given their dynamic and multifaceted role in energy metabolism, redox, and calcium balance, as well as cell death, mitochondria appear at the interface between environmental cues and the control of epigenetic identity. In this review, we describe how mitochondria have been implicated in the processes of acquisition and loss of stemness, with a specific focus on pluripotency. Dissecting the biological functions of mitochondria in stem cell homeostasis and differentiation will provide essential knowledge to understand the dynamics of cell fate modulation, and to establish improved stem cell‐based medical applications.     

Atrazine movement in soil: Comparison of field observations and PRZM simulations

The predictive capability of the pesticide root zone model (PRZM) was investigated for herbicide atrazine [2‐chloro‐4‐(ethylamino)‐6‐(isopropylamino)‐s‐triazine] in corn production under no‐till (NT) and conventional‐till (CT) management practices. Simulation values of atrazine residues obtained using our site‐specific soil and environmental data were compared with the actual values measured in soil samples taken from the root zones of the NT and CT plots during three growing seasons: 1986, 1987, and 1988. The mean concentration of atrazine in soil at each sampling time and depth after application, for each tillage treatment plot (NT or CT), was estimated based on the type of distribution (i.e., normal or lognormal). Overall, the PRZMs simulated concentrations for the top 10 cm of soil compared well with the atrazine residues measured in the CT plots, but overestimated measurements in NT plots. For example, in 1986 the mean atrazine concentration measured in soil samples taken 6 d after application from the top 10 cm of CT plots was 548 μg/kg (S.E. 198 μg/kg), and the PRZM predicted value was 690 μg/kg. In contrast, the mean atrazine concentration for the same soil depth increment in NT plots was 385 μg/kg (S.E. 154 μg/kg), with a PRZM predicted value of 674 μg/kg. Although the PRZM prediction was closer to the measured mean for atrazine concentrations in the top 10 cm of the CT system, the model did not transport atrazine to the lower soil depths, as the actual values have indicated in all 3 years. The results of this model comparison, especially for the lower soil depths (20 to 30 cm) in the NT practice, indicated that the PRZM model does not account for the preferential transport of, and, consequently, underestimates the atrazine residue levels in the lower soil profile under NT management systems.     

Patterns of roost site selection and use by Southern Ground-Hornbills in north-eastern South Africa

Different habitats may be used for the needs of various aspects of an animal’s life. Southern Ground-Hornbill Bucorvus leadbeateri groups announce their presence within year-round territories by calling at dawn from their overnight roost sites. Knowledge on ground-hornbill roosting habits is limited. Groups roost in large trees, apparently close to where they end up after daily foraging. We investigated patterns of roost site selection and use for four Southern Ground-Hornbill groups in the Associated Private Nature Reserves, north-eastern South Africa, based on data from GPS-satellite transmitters. The number of roost sites used per month averaged 15.4 ± 4.7 across all groups, indicating little evidence of strong preferences for specific sites. This number was least when groups were breeding, decreasing throughout the early wet season (October–December) and was lowest during the late wet season (January–March) when actively breeding groups frequently roosted close to the nest (54–83% of roosts <1 000 m of the nest). As might be expected, the mean monthly number of nights per roost peaked during the breeding season (December–January). Riparian habitats were preferred for roosting during the breeding season, whereas disturbed areas, as well as Combretumand mopane-dominated habitats were preferred during the dry non-breeding season. Adequate large trees not only for nesting, but also for roosting, particularly in riparian habitats, may therefore be an important and potentially limiting factor for the successful reproduction of Southern Ground-Hornbills.