A truncated progesterone receptor (PR-M) localizes to the mitochondrion and controls cellular respiration.

The cDNA for a novel truncated progesterone receptor (PR-M) was previously cloned from human adipose and aortic cDNA libraries. The predicted protein sequence contains 16 unique N-terminal amino acids, encoded by a sequence in the distal third intron of the progesterone receptor PR gene, followed by the same amino acid sequence encoded by exons 4 through 8 of the nuclear PR. Thus, PR-M lacks the N terminus A/B domains and the C domain for DNA binding, whereas containing the hinge and hormone-binding domains. In this report, we have localized PR-M to mitochondria using immunofluorescent localization of a PR-M-green fluorescent protein (GFP) fusion protein and in Western blot analyses of purified human heart mitochondrial protein. Removal of the putative N-terminal mitochondrial localization signal obviated association of PR-M with mitochondria, whereas addition of the mitochondrial localization signal to green fluorescent protein resulted in mitochondrial localization. Immunoelectron microscopy and Western blot analysis after mitochondrial fractionation identified PR-M in the outer mitochondrial membrane. Antibody specificity was shown by mass spectrometry identification of a PR peptide in a mitochondrial membrane protein isolation. Cell models of overexpression and gene silencing of PR-M demonstrated a progestin-induced increase in mitochondrial membrane potential and an increase in oxygen consumption consistent with an increase in cellular respiration. This is the first example of a truncated steroid receptor, lacking a DNA-binding domain that localizes to the mitochondrion and initiates direct non-nuclear progesterone action. We hypothesize that progesterone may directly affect cellular energy production to meet the increased metabolic demands of pregnancy.

Evaluation of respiratory function during Reeves stretcher use.

OBJECTIVE: We were aware of a small number of cases in our EMS system where patients in respiratory distress had a worsening of their condition after being removed from the home on a Reeves stretcher (RS). We sought to determine if this prehospital lifting device causes additional respiratory effort used in normal subjects by describing changes in heart rate, pulse oximetry, tidal volume, minute ventilation, and respiratory rate. METHODS: Forty-nine subjects were entered into this study. Data were collected while the subject was supine on the floor in the RS and once while suspended over the floor in the device. A randomized crossover design was used. Ten subjects were excluded because of inadvertent omission of a nose plug during spirometry. Data points were recorded in the final minute of a 3-minute exposure. Three minutes was chosen to simulate a prehospital transport time from the scene to the ambulance. Minute ventilation, tidal volume, heart rate, pulse oximetry, and respiratory rate were recorded for each subject during each phase. Subjects were also asked to rate the difficulty of breathing using the modified Borg scale. RESULTS: Data were obtained for 39 subjects. The mean respiratory rate while suspended was 9.9 +/- 3.0 breaths per minute compared to 9.1 +/- 2.5 breaths per minutes supine on the floor (p = 0.007). The mean minute ventilation while suspended in a RS was 8.17 +/- 3.25 L/min versus 7.37 +/- 2.37 while lying flat (p = 0.03). There was no difference in tidal volume, heart rate, pulse oximetry, or subjective modified Borg scale ratings. CONCLUSIONS: Subjects suspended in a RS for 3 minutes had statistically higher respiratory rates and minute ventilation than the same subjects lying flat. Although these modest changes are clinically insignificant in normal subjects, they could present a significant challenge to subjects in respiratory distress.

Systemic antibacterial activity of novel synthetic cyclic peptides.

Cyclic peptides with an even number of alternating d,l-alpha-amino acid residues are known to self-assemble into organic nanotubes. Such peptides previously have been shown to be stable upon protease treatment, membrane active, and bactericidal and to exert antimicrobial activity against Staphylococcus aureus and other gram-positive bacteria. The present report describes the in vitro and in vivo pharmacology of selected members of this cyclic peptide family. The intravenous (i.v.) efficacy of six compounds with MICs of less than 12 microg/ml was tested in peritonitis and neutropenic-mouse thigh infection models. Four of the six peptides were efficacious in vivo, with 50% effective doses in the peritonitis model ranging between 4.0 and 6.7 mg/kg against methicillin-sensitive S. aureus (MSSA). In the thigh infection model, the four peptides reduced the bacterial load 2.1 to 3.0 log units following administration of an 8-mg/kg i.v. dose. Activity against methicillin-resistant S. aureus was similar to MSSA. The murine pharmacokinetic profile of each compound was determined following i.v. bolus injection. Interestingly, those compounds with poor efficacy in vivo displayed a significantly lower maximum concentration of the drug in serum and a higher volume of distribution at steady state than compounds with good therapeutic properties. S. aureus was unable to easily develop spontaneous resistance upon prolonged exposure to the peptides at sublethal concentrations, in agreement with the proposed interaction with multiple components of the bacterial membrane canopy. Although additional structure-activity relationship studies are required to improve the therapeutic window of this class of antimicrobial peptides, our results suggest that these amphipathic cyclic d,l-alpha-peptides have potential for systemic administration and treatment of otherwise antibiotic-resistant infections.